Allosteric chromenone inhibitors of phosphoinositide 3-kinase (PI3K) for the treatment of diseases

Abstract

To provide allosteric chromenone inhibitors of phosphoinositide 3-kinase (PI3K) for the treatment of disease.SOLUTION: The present invention relates to a compound A, pharmaceutically acceptable salts thereof, production methods, and use thereof for the treatment of disease.SELECTED DRAWING: None

Description

Technical Field

[0001] Cross - reference to Related Applications This application claims priority to U.S. Provisional Application No. 63 / 421,636, filed on November 2, 2022 (Attorney Docket No. 30219_US_PRI); and U.S. Provisional Application No. 63 / 423,879, filed on November 9, 2022 (Attorney Docket No. 30219A_US_PRI), the contents of each of which are hereby incorporated by reference in their entirety.

[0002] The present invention relates to 2 - [[(1R)-1-(3,6 - dimethyl - 4 - oxo - 2 - phenyl - chromen - 8 - yl)ethyl]amino]benzoic acid (「Compound A」), its pharmaceutically acceptable salts, and its use for the treatment of diseases.

Background Art

[0003] The PIK3CA gene, which encodes the PI3K catalytic isoform p110α, is the most frequently mutated gene in solid tumors and is also the most frequent site of genetic changes within the PI3K pathway. PIK3CA mutations are most frequently seen in endometrial cancer, breast cancer, and head and neck cancer. Approximately 40% of patients with HR+ / HER2 - breast cancer have activating mutations in PIK3CA that activate the p110α and PI3K / AKT / mTOR signaling network. H1047R is the most common missense mutation in PIK3CA.

[0004] Several PI3K targeting agents are being tested in breast cancer patients. The PI3Kα-specific inhibitor alpelisib, in combination with fulvestrant, in the randomized phase 3 trial (SOLAR-1; NCT02437318), increased the progression-free survival to 11.0 months compared to 5.7 months with fulvestrant alone, leading to the FDA approval of alpelisib in combination with fulvestrant for the treatment of HR+ / HER2- PIK3CA-mutant advanced or metastatic breast cancer patients. This approval represented a significant therapeutic advance for PIK3CA mutant breast cancer, but alpelisib, along with other investigational PI3Kα inhibitors in the clinic, inhibits both wild-type (WT) and mutant PI3Kα with nearly equal potency. As a result, their efficacy is potentially limited by on-target WT PI3Kα-mediated toxicities including dose-limiting hyperglycemia and skin and GI toxicities, which have somewhat restricted the broad clinical utility of PI3Kα inhibitors.

[0005] There is a need for PI3Kα inhibitors with improved therapeutic indices for the treatment of diseases associated with mutant PI3K, including PIK3CA mutant cancers. Solid forms of PI3Kα inhibitors with advantageous physical stability, chemical stability, solubility, or pharmacokinetic properties are also needed. SUMMARY OF THE INVENTION

[0006] In one aspect, a solid form of Compound A is provided.

[0007] In another aspect, a pharmaceutically acceptable salt of Compound A, and solid forms thereof are provided.

[0008] In another aspect, a therapy comprising Compound A, or a pharmaceutically acceptable salt thereof, for the treatment of diseases such as PIK3CA mutant cancers is provided. BRIEF DESCRIPTION OF THE DRAWINGS

[0009]

Figure 1

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Mode for Carrying Out the Invention

[0010] Compound 2-[[(1R)-1-(3,6-dimethyl-4-oxo-2-phenyl-chromen-8-yl)ethyl]amino]benzoic acid (“Compound A”) is a potent and mutation-selective inhibitor of PI3Kα H1047R.

[0011]

Chemical Formula

[0012] In one aspect, solid forms of Compound A are provided herein.

[0013] Compound A, Form A In one aspect, there is provided crystalline 2-[[(1R)-1-(3,6-dimethyl-4-oxo-2-phenyl-chromen-8-yl)ethyl]amino]benzoic acid, also referred to as Compound A Form A. In one embodiment, Compound A Form A is characterized by an X-ray powder diffraction pattern using CuKα radiation having at least one peak at diffraction angles 2θ selected from 7.8° ± 0.2°, 12.1° ± 0.2°, 13.7° ± 0.2°, 14.1° ± 0.2°, 16.8° ± 0.2°, 17.5° ± 0.2°, 18.2° ± 0.2°, 18.9° ± 0.2°, 19.5° ± 0.2°, 20.7° ± 0.2°, 21.2° ± 0.2°, and 24.1° ± 0.2°. In another embodiment, Compound A Form A is characterized by an X-ray powder diffraction pattern using CuKα radiation having a peak at a diffraction angle 2θ of 12.1° ± 0.2° together with at least one peak selected from 14.1° ± 0.2°, 16.8° ± 0.2°, 18.9° ± 0.2°, and 20.7° ± 0.2°. In another embodiment, Compound A Form A is characterized by an X-ray powder diffraction pattern using CuKα radiation having a peak at a diffraction angle 2θ of 12.1° ± 0.2° together with at least two peaks selected from 14.1° ± 0.2°, 16.8° ± 0.2°, 18.9° ± 0.2°, and 20.7° ± 0.2°. In another embodiment, Compound A Form A is characterized by an X-ray powder diffraction pattern using CuKα radiation having a peak at a diffraction angle 2θ of 12.1° ± 0.2° together with at least three peaks selected from 14.1° ± 0.2°, 16.8° ± 0.2°, 18.9° ± 0.2°, and 20.7° ± 0.2°. In another embodiment, Compound A Form A is characterized by an X-ray powder diffraction pattern using CuKα radiation having a peak at a diffraction angle 2θ of 12.1° ± 0.2° together with peaks at 14.1° ± 0.2°, 16.8° ± 0.2°, 18.9° ± 0.2°, and 20.7° ± 0.2°.In another embodiment, Compound A Form A is characterized by an X-ray powder diffraction pattern using CuKa radiation having peaks at diffraction angles 2 theta of 7.8° ± 0.2°, 12.1° ± 0.2°, 13.7° ± 0.2°, 14.1° ± 0.2°, 16.8° ± 0.2°, 17.5° ± 0.2°, 18.2° ± 0.2°, 18.9° ± 0.2°, 19.5° ± 0.2°, 20.7° ± 0.2°, 21.2° ± 0.2° and 24.1° ± 0.2°.

[0014] Compound A Form B In another aspect, there is provided crystalline 2-[[(1R)-1-(3,6-dimethyl-4-oxo-2-phenyl-chromen-8-yl)ethyl]amino]benzoic acid Form B, also referred to as Compound A Form B. In one embodiment, Compound A Form B is characterized by an X-ray powder diffraction pattern using CuKa radiation having at least one peak at a diffraction angle 2 theta selected from 7.0° ± 0.2°, 9.7° ± 0.2°, 11.9° ± 0.2°, 14.9° ± 0.2°, and 17.4° ± 0.2°. In another embodiment, Compound A Form B is characterized by an X-ray powder diffraction pattern using CuKa radiation having a peak at a diffraction angle 2 theta of 9.7° ± 0.2° together with at least one peak selected from 14.9° ± 0.2°, 11.9° ± 0.2°, 17.4° ± 0.2° and 7.0° ± 0.2°. In another embodiment, Compound A Form B is characterized by an X-ray powder diffraction pattern using CuKa radiation having a peak at a diffraction angle 2 theta of 9.7° ± 0.2° together with at least two peaks selected from 14.9° ± 0.2°, 11.9° ± 0.2°, 17.4° ± 0.2° and 7.0° ± 0.2°. In another embodiment, Compound A Form B is characterized by an X-ray powder diffraction pattern using CuKa radiation having a peak at a diffraction angle 2 theta of 9.7° ± 0.2° together with at least three peaks selected from 14.9° ± 0.2°, 11.9° ± 0.2°, 17.4° ± 0.2° and 7.0° ± 0.2°. In another embodiment, Compound A Form B is characterized by an X-ray powder diffraction pattern using CuKa radiation having peaks at diffraction angles 2 theta of 7.0° ± 0.2°, 9.7° ± 0.2°, 11.9° ± 0.2°, 14.9° ± 0.2° and 17.4° ± 0.2°.

[0015] Form C of Compound A In another aspect, crystalline 2-[[(1R)-1-(3,6-dimethyl-4-oxo-2-phenyl-chromen-8-yl)ethyl]amino]benzoic acid, also referred to as Form C of Compound A, is provided. In one embodiment, Compound A, Form C, is characterized by an X-ray powder diffraction pattern using CuKα radiation having at least one peak at diffraction angles 2θ selected from 7.4° ± 0.2°, 8.5° ± 0.2°, 10.6° ± 0.2°, 13.4° ± 0.2° and 15.7° ± 0.2°. In another embodiment, Compound A, Form C, is characterized by an X-ray powder diffraction pattern using CuKα radiation having a peak at a diffraction angle 2θ of 13.4° ± 0.2° together with at least one peak selected from 8.5° ± 0.2°, 15.7° ± 0.2°, 10.6° ± 0.2° and 7.4° ± 0.2°. In another embodiment, Compound A, Form C, is characterized by an X-ray powder diffraction pattern using CuKα radiation having a peak at a diffraction angle 2θ of 13.4° ± 0.2° together with at least two peaks selected from 8.5° ± 0.2°, 15.7° ± 0.2°, 10.6° ± 0.2° and 7.4° ± 0.2°. In another embodiment, Compound A, Form C, is characterized by an X-ray powder diffraction pattern using CuKα radiation having a peak at a diffraction angle 2θ of 13.4° ± 0.2° together with at least three peaks selected from 8.5° ± 0.2°, 15.7° ± 0.2°, 10.6° ± 0.2° and 7.4° ± 0.2°. In another embodiment, Compound A, Form C, is characterized by an X-ray powder diffraction pattern using CuKα radiation having peaks at diffraction angles 2θ of 7.4° ± 0.2°, 8.5° ± 0.2°, 10.6° ± 0.2°, 13.4° ± 0.2° and 15.7° ± 0.2°.

[0016] In another aspect, the tromethamine salt of 2-[[(1R)-1-(3,6-dimethyl-4-oxo-2-phenyl-chromen-8-yl)ethyl]amino]benzoic acid is provided herein. Particular tromethamine salts of Compound A may have advantageous physical stability, chemical stability, solubility, or pharmacokinetic properties. Particular tromethamine salts of Compound A may have processability or other manufacturing advantages. Particular tromethamine salts of Compound A may provide chiral enhancement of Compound A upon crystallization of the tromethamine salt.

[0017] Compound A Tromethamine Salt Form A In another aspect, there is provided a crystalline tromethamine salt of 2-[[(1R)-1-(3,6-dimethyl-4-oxo-2-phenyl-chromen-8-yl)ethyl]amino]benzoic acid, referred to as Compound A tromethamine salt Form A. In one embodiment, Compound A tromethamine salt Form A is characterized by an X-ray powder diffraction pattern using CuKα radiation having at least one peak at a diffraction angle 2θ selected from 6.4° ± 0.2°, 8.4° ± 0.2°, 10.9° ± 0.2°, 11.8° ± 0.2°, 13.0° ± 0.2°, 16.5° ± 0.2°, 16.9° ± 0.2°, 22.1° ± 0.2°, 23.0° ± 0.2°, and 24.9° ± 0.2°. In another embodiment, Compound A tromethamine salt Form A is characterized by an X-ray powder diffraction pattern using CuKα radiation having a peak at a diffraction angle 2θ of 6.4° ± 0.2° together with at least one peak selected from 8.4° ± 0.2°, 10.9° ± 0.2°, 16.9° ± 0.2°, and 22.1° ± 0.2°. In another embodiment, Compound A tromethamine salt Form A is characterized by an X-ray powder diffraction pattern using CuKα radiation having a peak at a diffraction angle 2θ of 6.4° ± 0.2° together with at least two peaks selected from 8.4° ± 0.2°, 10.9° ± 0.2°, 16.9° ± 0.2°, and 22.1° ± 0.2°. In another embodiment, Compound A tromethamine salt Form A is characterized by an X-ray powder diffraction pattern using CuKα radiation having a peak at a diffraction angle 2θ of 6.4° ± 0.2° together with at least three peaks selected from 8.4° ± 0.2°, 10.9° ± 0.2°, 16.9° ± 0.2°, and 22.1° ± 0.2°. In another embodiment, Compound A tromethamine salt Form A is characterized by an X-ray powder diffraction pattern using CuKα radiation having a peak at a diffraction angle 2θ of 6.4° ± 0.2° together with peaks at 8.4° ± 0.2°, 10.9° ± 0.2°, 16.9° ± 0.2°, and 22.1° ± 0.2°.In another embodiment, Compound A tromethamine salt Form A is characterized by an X-ray powder diffraction pattern using CuKa radiation having peaks at diffraction angles 2 theta of 6.4° ± 0.2°, 8.4° ± 0.2°, 10.9° ± 0.2°, 11.8° ± 0.2°, 13.0° ± 0.2°, 16.5° ± 0.2°, 16.9° ± 0.2°, 22.1° ± 0.2°, 23.0° ± 0.2° and 24.9° ± 0.2°.

[0018] In another embodiment, Compound A tromethamine salt Form A comprises at least one peak referenced to glycine (external reference at 176.5 ppm) selected from 179.0, 158.7, 151.7, 149.7, 136.3, 134.7, 132.9, 129.3, 127.4, 125.2, 121.7, 117.0, 115.5, 115.2, 110.4, 64.1, 63.2, 45.3, 22.6, 20.3, and 11.6 ppm (± 0.2 ppm each). 13 characterized by a 13C solid state NMR (100.6 MHz) spectrum. In another embodiment, Compound A tromethamine salt Form A comprises at least one peak referenced to glycine (external reference at 176.5 ppm) selected from 179.0, 129.3, 63.2, 20.3 and 11.6 ppm (± 0.2 ppm each). 13 characterized by a 13C solid state NMR (100.6 MHz) spectrum. In another embodiment, Compound A tromethamine salt Form A comprises peaks referenced to glycine (external reference at 176.5 ppm) at 179.0, 158.7, 151.7, 149.7, 136.3, 134.7, 132.9, 129.3, 127.4, 125.2, 121.7, 117.0, 115.5, 115.2, 110.4, 64.1, 63.2, 45.3, 22.6, 20.3 and 11.6 ppm (± 0.2 ppm each). 13 characterized by a 13C solid state NMR (100.6 MHz) spectrum.

[0019] Compound A tromethamine salt Form C In another aspect, there is provided a crystalline tromethamine salt of 2-[[(1R)-1-(3,6-dimethyl-4-oxo-2-phenyl-chromen-8-yl)ethyl]amino]benzoic acid, referred to as Compound A tromethamine salt Form C. In one embodiment, Compound A tromethamine salt Form C is characterized by an X-ray powder diffraction pattern using CuKα radiation having at least one peak at diffraction angles 2θ selected from 10.6° ± 0.2°, 13.2° ± 0.2°, 14.5° ± 0.2°, 15.9° ± 0.2° and 17.4° ± 0.2°. In another embodiment, Compound A tromethamine salt Form C is characterized by an X-ray powder diffraction pattern using CuKα radiation having a peak at a diffraction angle 2θ of 15.9° ± 0.2° together with at least one peak selected from 10.6° ± 0.2°, 17.4° ± 0.2°, 13.2° ± 0.2° and 14.5° ± 0.2°. In another embodiment, Compound A tromethamine salt Form C is characterized by an X-ray powder diffraction pattern using CuKα radiation having a peak at a diffraction angle 2θ of 15.9° ± 0.2° together with at least two peaks selected from 10.6° ± 0.2°, 17.4° ± 0.2°, 13.2° ± 0.2° and 14.5° ± 0.2°. In another embodiment, Compound A tromethamine salt Form C is characterized by an X-ray powder diffraction pattern using CuKα radiation having a peak at a diffraction angle 2θ of 15.9° ± 0.2° together with at least three peaks selected from 10.6° ± 0.2°, 17.4° ± 0.2°, 13.2° ± 0.2° and 14.5° ± 0.2°. In one embodiment, Compound A tromethamine salt Form C is characterized by an X-ray powder diffraction pattern using CuKα radiation having peaks at diffraction angles 2θ of 10.6° ± 0.2°, 13.2° ± 0.2°, 14.5° ± 0.2°, 15.9° ± 0.2° and 17.4° ± 0.2°.

[0020] Compound A tromethamine salt Form D In another aspect, provided is a crystalline tromethamine salt of 2-[[(1R)-1-(3,6-dimethyl-4-oxo-2-phenyl-chromen-8-yl)ethyl]amino]benzoic acid, referred to as Compound A tromethamine salt Form D. In one embodiment, Compound A tromethamine salt Form D is characterized by an X-ray powder diffraction pattern using CuKα radiation having at least one peak at a diffraction angle 2θ selected from 6.3° ± 0.2°, 11.1° ± 0.2°, 12.6° ± 0.2°, 17.1° ± 0.2°, and 18.9° ± 0.2°. In another embodiment, Compound A tromethamine salt Form D is characterized by an X-ray powder diffraction pattern using CuKα radiation having a peak at a diffraction angle 2θ of 11.1° ± 0.2° together with at least one peak selected from 12.6° ± 0.2°, 17.1° ± 0.2°, 6.3° ± 0.2° and 18.9° ± 0.2°. In another embodiment, Compound A tromethamine salt Form D is characterized by an X-ray powder diffraction pattern using CuKα radiation having a peak at a diffraction angle 2θ of 11.1° ± 0.2° together with at least two peaks selected from 12.6° ± 0.2°, 17.1° ± 0.2°, 6.3° ± 0.2° and 18.9° ± 0.2°. In another embodiment, Compound A tromethamine salt Form D is characterized by an X-ray powder diffraction pattern using CuKα radiation having a peak at a diffraction angle 2θ of 11.1° ± 0.2° together with at least three peaks selected from 12.6° ± 0.2°, 17.1° ± 0.2°, 6.3° ± 0.2° and 18.9° ± 0.2°. In one embodiment, Compound A tromethamine salt Form D is characterized by an X-ray powder diffraction pattern using CuKα radiation having peaks at diffraction angles 2θ of 6.3° ± 0.2°, 11.1° ± 0.2°, 12.6° ± 0.2°, 17.1° ± 0.2°, and 18.9° ± 0.2°.

[0021] In another aspect, provided herein is an erbumine salt of 2-[[(1R)-1-(3,6-dimethyl-4-oxo-2-phenyl-chromen-8-yl)ethyl]amino]benzoic acid. Certain erbumine salts of Compound A may have advantageous physical stability, chemical stability, solubility, or pharmacokinetic properties. Certain erbumine salts of Compound A may have processability or other manufacturing advantages.

[0022] Albumin salt form A of Compound A In another aspect, there is provided a crystalline albumin salt of 2-[[(1R)-1-(3,6-dimethyl-4-oxo-2-phenyl-chromen-8-yl)ethyl]amino]benzoic acid, referred to as Compound A albumin salt Form A. In one embodiment, Compound A albumin salt Form A is characterized by an X-ray powder diffraction pattern using CuKα radiation having at least one peak at diffraction angles 2θ selected from 6.5° ± 0.2°, 10.5° ± 0.2°, 11.1° ± 0.2°, 15.2° ± 0.2°, 15.9° ± 0.2°, 17.6° ± 0.2°, 18.0° ± 0.2°, 19.3° ± 0.2°, 21.5° ± 0.2°, 22.2° ± 0.2°, 22.7° ± 0.2°, and 26.3° ± 0.2°. In another embodiment, Compound A albumin salt Form A is characterized by an X-ray powder diffraction pattern using CuKα radiation having a peak at a diffraction angle 2θ of 11.1° ± 0.2° together with at least one peak selected from 10.5° ± 0.2°, 15.2° ± 0.2°, 18.0° ± 0.2°, and 19.3° ± 0.2°. In another embodiment, Compound A albumin salt Form A is characterized by an X-ray powder diffraction pattern using CuKα radiation having a peak at a diffraction angle 2θ of 11.1° ± 0.2° together with at least two peaks selected from 10.5° ± 0.2°, 15.2° ± 0.2°, 18.0° ± 0.2°, and 19.3° ± 0.2°. In another embodiment, Compound A albumin salt Form A is characterized by an X-ray powder diffraction pattern using CuKα radiation having a peak at a diffraction angle 2θ of 11.1° ± 0.2° together with at least three peaks selected from 10.5° ± 0.2°, 15.2° ± 0.2°, 18.0° ± 0.2°, and 19.3° ± 0.2°. In another embodiment, Compound A albumin salt Form A is characterized by an X-ray powder diffraction pattern using CuKα radiation having a peak at a diffraction angle 2θ of 11.1° ± 0.2° together with peaks at 10.5° ± 0.2°, 15.2° ± 0.2°, 18.0° ± 0.2°, and 19.3° ± 0.2°.In one embodiment, compound A albumin salt form A is characterized by an X-ray powder diffraction pattern using CuKa radiation having peaks at diffraction angles 2 theta of 66.5° ± 0.2°, 10.5° ± 0.2°, 11.1° ± 0.2°, 15.2° ± 0.2°, 15.9° ± 0.2°, 17.6° ± 0.2°, 18.0° ± 0.2°, 19.3° ± 0.2°, 21.5° ± 0.2°, 22.2° ± 0.2°, 22.7° ± 0.2°, and 26.3° ± 0.2°.

[0023] In another embodiment, compound A albumin salt form A comprises at least one peak referenced to glycine (external reference at 176.5 ppm) selected from 177.4, 174.8, 159.8, 151.7, 149.5, 134.0, 132.6, 130.7, 130.3, 129.0, 128.1, 123.1, 122.4, 119.4, 116.8, 115.9, 112.3, 53.0, 47.5, 27.4, 23.3, 21.3, and 11.3 ppm (each ± 0.2 ppm). 13 characterized by a 13C solid state NMR (100.6 MHz) spectrum. In another embodiment, compound A, albumin salt form A, comprises at least one peak referenced to glycine (external reference at 176.5 ppm) selected from 177.4, 132.6, 27.4, 21.3 and 11.3 ppm (each ± 0.2 ppm). 13 characterized by a 13C solid state NMR (100.6 MHz) spectrum. In another embodiment, compound A, albumin salt form A, comprises peaks referenced to glycine (external reference at 176.5 ppm) at 177.4, 174.8, 159.8, 151.7, 149.5, 134.0, 132.6, 130.7, 130.3, 129.0, 128.1, 123.1, 122.4, 119.4, 116.8, 115.9, 112.3, 53.0, 47.5, 27.4, 23.3, 21.3 and 11.3 ppm (each ± 0.2 ppm). 13 characterized by a 13C solid state NMR (100.6 MHz) spectrum.

[0024] Therapeutic use Also provided herein is a treatment method comprising compound A, or a pharmaceutically acceptable salt thereof, for treating patients having a disease including PIK3CA mutant cancer, such as PIK3CA mutant, advanced or metastatic breast cancer, or other solid tumors having a PIK3CA mutation. Compound A, or a pharmaceutically acceptable salt thereof, can be used in monotherapy or in combination with one or more additional therapeutic agents. The therapy can provide new treatment options for patients and, in some patients, can provide enhanced and / or unexpected beneficial therapeutic effects compared to known therapies.

[0025] The effectiveness of cancer treatment can be measured by various endpoints commonly used in the evaluation of cancer treatment (including, but not limited to, tumor regression, reduction in tumor weight or size, time to progression, overall survival, progression-free survival, overall response rate, duration of response, best overall response, disease control rate, clinical benefit rate, time to response, and quality of life). The therapeutic agent can cause inhibition of metastatic spread without accompanying shrinkage of the primary tumor, can induce shrinkage of the primary tumor, or can simply exert a tumor-suppressive effect. For example, a novel approach can optionally be used to determine the effectiveness of any particular monotherapy or combination therapy of the present invention, including measurement of plasma or urine markers of angiogenesis and / or cell cycle activity, measurement of tissue-based biomarkers for angiogenesis and / or cell cycle activity, and measurement of response by radiographic imaging.

[0026] In one aspect, provided is a method for treating a patient having a disease associated with mutant phosphoinositide 3-kinase (PI3K), comprising administering to the patient an effective amount of compound A, or a pharmaceutically acceptable salt thereof.

[0027] In another aspect, provided is a method for treating a patient having PIK3CA mutant cancer, comprising administering to the patient an effective amount of compound A, or a pharmaceutically acceptable salt thereof.

[0028] In another aspect, there is provided a method of treating a patient having a PIK3CA-mutated solid tumor, comprising administering to the patient an effective amount of Compound A, or a pharmaceutically acceptable salt thereof.

[0029] In another aspect, there is provided a method of treating a patient having a PIK3CA-mutated breast cancer, comprising administering to the patient an effective amount of Compound A, or a pharmaceutically acceptable salt thereof.

[0030] In another aspect, there is provided a method of treating a patient having a PIK3CA-mutated, advanced or metastatic breast cancer, comprising administering to the patient an effective amount of Compound A, or a pharmaceutically acceptable salt thereof.

[0031] In another aspect, there is provided a method of treating a patient having CLOVES syndrome (Congenital Lipomatous Overgrowth, Vascular Malformations, Epidermal Nevus, Scoliosis / Skeletal, and Spinal Cord Syndrome) or PIK3CA-related overgrowth syndrome (PROS), comprising administering to the patient an effective amount of Compound A, or a pharmaceutically acceptable salt thereof.

[0032] In another aspect, there is provided Compound A, or a pharmaceutically acceptable salt thereof, for use in the treatment of a disease associated with mutant phosphoinositide 3-kinase (PI3K).

[0033] In another aspect, there is provided Compound A, or a pharmaceutically acceptable salt thereof, for use in the treatment of a PIK3CA-mutated cancer.

[0034] In another aspect, there is provided Compound A, or a pharmaceutically acceptable salt thereof, for use in the treatment of a PIK3CA-mutated solid tumor.

[0035] In another aspect, there is provided Compound A, or a pharmaceutically acceptable salt thereof, for use in the treatment of a PIK3CA-mutated breast cancer.

[0036] In another aspect, there is provided Compound A, or a pharmaceutically acceptable salt thereof, for use in the treatment of a PIK3CA-mutated, advanced or metastatic breast cancer.

[0037] In another aspect, compound A, or a pharmaceutically acceptable salt thereof, is provided for use in the treatment of CLOVES syndrome (Congenital Lipomatous Overgrowth, Vascular Malformations, Epidermal Nevi, Scoliosis / Skeletal, and Spinal Cord Syndrome) or PIK3CA-related overgrowth syndrome (PROS).

[0038] In another aspect, the use of compound A, or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for treating a disease associated with mutant phosphoinositide 3-kinase (PI3K) is provided.

[0039] In another aspect, the use of compound A, or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for treating PIK3CA mutant cancer is provided.

[0040] In another aspect, the use of compound A, or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for treating PIK3CA mutant solid tumors is provided.

[0041] In another aspect, the use of compound A, or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for treating PIK3CA mutant breast cancer is provided.

[0042] In another aspect, the use of compound A, or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for treating PIK3CA mutant, advanced or metastatic breast cancer is provided.

[0043] In another aspect, the use of compound A, or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for treating CLOVES syndrome (Congenital Lipomatous Overgrowth, Vascular Malformations, Epidermal Nevi, Scoliosis / Skeletal, and Spinal Cord Syndrome) or PIK3CA-related overgrowth syndrome (PROS) is provided.

[0044] In one embodiment, PIK3CA-mutated cancers include acute lymphoblastic leukemia (ALL), acute myeloid leukemia (AML), adrenocortical carcinoma, AIDS-related cancers, AIDS-related lymphomas, anal cancer, astrocytoma, basal cell carcinoma, cholangiocarcinoma, bladder cancer, bone cancer, osteosarcoma, malignant fibrous histiocytoma, brain tumors, breast cancer, bronchial tumors, Burkitt lymphoma, carcinoid tumors, cancer of unknown primary origin, cardiac (heart) tumors, atypical teratoid / rhabdoid tumors, primary CNS lymphoma, cervical cancer, cholangiocarcinoma, chordoma, chronic lymphocytic leukemia (CLL), chronic myeloid leukemia (CML), colorectal cancer, craniopharyngioma, cutaneous T-cell lymphoma, mycosis fungoides, Sézary syndrome, ductal carcinoma in situ (DCIS), germ cell tumors, medulloblastoma, endometrial cancer, ependymoma, esophageal cancer, neuroblastoma, Ewing sarcoma, extracranial germ cell tumors, extragonadal germ cell tumors, fallopian tube cancer, gallbladder cancer, gastric cancer, gastrointestinal carcinoid tumors, gastrointestinal stromal tumors (GIST), germ cell tumors, gestational trophoblastic disease, hairy cell leukemia, head and neck cancers, hepatocellular carcinoma, Langerhans cell histiocytosis, Hodgkin lymphoma, islet cell tumors, pancreatic neuroendocrine tumors, Kaposi sarcoma, kidney cancer, laryngeal cancer, leukemia, liver cancer, lung cancer, lymphomas, male breast cancer, intraocular melanoma, Merkel cell carcinoma, malignant mesothelioma, metastatic cancer, metastatic squamous neck cancer, midline cancer with NUT gene alteration, mouth cancer, multiple endocrine neoplasia syndromes, multiple myeloma / plasma cell tumors, myelodysplastic syndromes, myelodysplastic tumors, myeloproliferative tumors, chronic myeloproliferative tumors, nasal and paranasal cavity cancers, nasopharyngeal cancer, neuroblastoma, non-Hodgkin lymphoma, non-small cell lung cancer, oral cancer, lip cancer and oral cavitycancer), oral pharyngeal cancer, malignant fibrous histiocytoma of bone, ovarian cancer, pancreatic cancer, pancreatic neuroendocrine tumor (islet cell tumor), papillomatosis, paraganglioma, paranasal and nasal cavity cancer, parathyroid cancer, penile cancer, pharyngeal cancer, pheochromocytoma, pituitary tumor, plasmacytoma, multiple myeloma, pleuropulmonary blastoma, primary central nervous system (CNS) lymphoma, primary peritoneal cancer, prostate cancer, rectal cancer, recurrent cancer, renal cell (kidney) cancer, retinoblastoma, rhabdomyosarcoma, salivary gland cancer, sarcoma, pediatric hemangioma, skin cancer, small cell lung cancer, small intestine cancer, soft tissue sarcoma, squamous cell carcinoma of the skin, testicular cancer, oral pharyngeal cancer, hypopharyngeal cancer, thymoma, thymic carcinoma, thyroid cancer, tracheobronchial tumor, transitional cell carcinoma of the renal pelvis and ureter, urethral cancer, uterine sarcoma, vaginal cancer, hemangioma, vulvar cancer, and Wilms tumor.

[0045] In one embodiment, PIK3CA mutant cancer is endometrial cancer, breast cancer, esophageal squamous cell carcinoma, cervical squamous cell carcinoma, cervical adenocarcinoma, colorectal adenocarcinoma, bladder urothelial carcinoma, glioblastoma, ovarian cancer, non-small cell lung cancer, esophagogastric cancer, schwannoma, head and neck squamous cell carcinoma, melanoma, esophagogastric adenocarcinoma, soft tissue sarcoma, prostate cancer, fibrolamellar carcinoma, hepatocellular carcinoma, diffuse glioma, colorectal cancer, pancreatic cancer, cholangiocarcinoma, B cell lymphoma, mesothelioma, adrenocortical carcinoma, non-clear cell type renal cell carcinoma, clear cell type renal cell carcinoma, germ cell carcinoma, thymic tumor, pheochromocytoma, variously mixed neuroepithelial tumors, thyroid cancer, leukemia, or encapsulated glioma.

[0046] In one embodiment, PIK3CA mutant cancer is breast cancer, brain cancer, prostate cancer, endometrial cancer, gastric cancer, leukemia, lymphoma, sarcoma, colorectal cancer, lung cancer, ovarian cancer, skin cancer, or head and neck cancer.

[0047] In one embodiment, PIK3CA mutant cancer is breast cancer, prostate cancer or brain cancer. In one embodiment, PIK3CA mutant cancer is breast cancer. In one embodiment, PIK3CA mutant cancer is prostate cancer. In one embodiment, PIK3CA mutant cancer is brain cancer.

[0048] In one embodiment, the PIK3CA mutant cancer is a breast neoplasm, a thyroid neoplasm, an ovarian neoplasm, a non-small cell lung cancer, an endometrial neoplasm, or a pancreatic neoplasm. In one embodiment, the PIK3CA mutant cancer is a breast neoplasm. In one embodiment, the PIK3CA mutant cancer is a thyroid neoplasm. In one embodiment, the PIK3CA mutant cancer is an ovarian neoplasm. In one embodiment, the PIK3CA mutant cancer is a non-small cell lung cancer. In one embodiment, the PIK3CA mutant cancer is an endometrial neoplasm. In one embodiment, the PIK3CA mutant cancer is a pancreatic neoplasm.

[0049] In one embodiment, the PIK3CA mutant, advanced or metastatic breast cancer is a PIK3CA H1047R mutant, advanced or metastatic breast cancer. In one embodiment, the PIK3CA mutant, advanced or metastatic breast cancer is a hormone receptor positive (HR+), human epidermal growth factor receptor 2 negative (HER2-), PIK3CA mutant, advanced or metastatic breast cancer. In one embodiment, the PIK3CA mutant, advanced or metastatic breast cancer is an estrogen receptor positive (ER+), human epidermal growth factor receptor 2 negative (HER2-), PIK3CA mutant, advanced or metastatic breast cancer. In one embodiment, the PIK3CA mutant, advanced or metastatic breast cancer is a hormone receptor positive (HR+), human epidermal growth factor receptor 2 negative (HER2-), PIK3CA H1047R mutant, advanced or metastatic breast cancer. In one embodiment, the PIK3CA mutant, advanced or metastatic breast cancer is an estrogen receptor positive (ER+), human epidermal growth factor receptor 2 negative (HER2-), PIK3CA H1047R mutant, advanced or metastatic breast cancer.

[0050] In one embodiment, the PIK3CA mutant solid tumor is a PIK3CA mutant advanced solid tumor. In one embodiment, the PIK3CA mutant advanced solid tumor is selected from gynecological cancers, head and neck cancers, and triple negative breast cancer. In one embodiment, the PIK3CA mutant advanced solid tumor is a gynecological cancer. In one embodiment, the PIK3CA mutant advanced solid tumor is a head and neck cancer. In one embodiment, the PIK3CA mutant advanced solid tumor is a triple negative breast cancer.

[0051] Pharmaceutical composition Compound A, or a pharmaceutically acceptable salt thereof, can be formulated for oral administration in the form of tablets, capsules (each of which includes sustained release or extended release formulations), pills, powders, granules, elixirs, tinctures, suspensions, syrups, and emulsions, etc. Compound A, or a pharmaceutically acceptable salt thereof, can also be formulated for intravenous (bolus or infusion), intraperitoneal, topical, subcutaneous, intramuscular, or transdermal (e.g., patch) administration using all forms well known to those skilled in the pharmaceutical art.

[0052] Compound A, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof, can be administered to a subject by any convenient route of administration, whether systemically / peripherally or locally (i.e., at the desired site of action).

[0053] Routes of administration include oral (e.g., by ingestion); buccal; sublingual; transdermal (including those by patch, plaster, etc.); transmucosal (including those by patch, plaster, etc.); intranasal (e.g., by nasal spray); ocular (e.g., by eye drops); pulmonary (e.g., via aerosol, e.g., used through the mouth or nose, e.g., by inhalation or insufflation therapy); rectal (e.g., by suppository or enema); vaginal (e.g., by pessary); parenteral by injection, including subcutaneous, intradermal, intramuscular, intravenous, intra - arterial, intracardiac, intrathecal, intraspinal, intracapsular, subcapsular, intraorbital, intraperitoneal, intratracheal, subepidermal, intra - articular, subarachnoid, and intrasternal, etc.; depot or reservoir, e.g., by implantation subcutaneously or intramuscularly; and the like, but are not limited thereto.

[0054] Synthetic Method Compound A can be synthesized using the methods described below, along with synthetic methods known in the field of organic synthetic chemistry or modifications thereof understood by those skilled in the art. Preferred methods include, but are not limited to, the methods described below. Compound A can be synthesized according to the steps outlined in General Schemes 1 and 2. The starting materials are either commercially available or prepared by known procedures in the reported literature or as illustrated below.

[0055]

Chem.

[0056] Scheme 1 shows an exemplary preparation of compound A. By acylation of the substituted phenol (1), the ester (2) can be obtained. The ester (2) can undergo rearrangement to the hydroxyaryl ketone (3) under Lewis acid (e.g., AlCl3) or Brønsted acid (e.g., triflic acid) conditions. Acidic condensation of the aryl aldehyde with the hydroxyaryl ketone (3) can give the keto-alkene (4) which can cyclize to the 2-substituted chromen-4-one (5). Alternatively, alkylation of the hydroxyaryl ketone (3) with an aryl halide in the presence of a base (e.g., pyridine or lithium bis(trimethylsilyl)amide), followed by acidic conditions (e.g., HCl) can affect cyclization to the 2-substituted chromen-4-one (5).

[0057] Phenyl bromide (5) can be acylated by palladium catalysis to produce acylchromen-4-one (6). Exemplary palladium catalyst conditions include phenyl bromide (5) in about 30 - 35 equivalents of dioxane at 95 °C for about 16 hours, about 5 - 10 mol% of PdCl2(Ph3)2, and about 1.2 mol% of tributyl(1-ethoxyvinyl)stannane; or phenyl bromide (5), about 1 mol% of Pd(OAc)2, about 2 mol% of 1,3-bis(diphenylphosphino)propane, about 5 equivalents of butyl vinyl ether, about 3 equivalents of trimethylamine, and about 10 volumes of ethylene glycol at about 100 °C for about 16 hours. A ketimine (7) can be obtained by condensation of the ketone (6) with tert-butanesulfinamide using a Lewis acid dehydrating agent such as titanium(IV) alkoxide. The asymmetric reduction of the sulfinimine (7) can be affected by a borohydride reagent in the presence of a transition metal catalyst such as cerium(III) chloride to produce a chiral-rich sulfinamide (8). The sulfinamide (8) can be converted to benzylamine (9) using removal of the sulfinyl group under acidic conditions, and this can be alkylated with an aryl halide (10) under Finkelstein or Ullmann-type conditions to obtain compound A.

[0058]

Chemical Structure

[0059] Scheme 2 shows another exemplary preparation of compound A. Ketone (6) can be reduced to hydroxy compound (11) using a chiral catalyst such as a Noyori catalyst. The hydroxyl group can be converted to a leaving group with methanesulfonic anhydride or methanesulfonyl chloride to obtain mesylate (12). Mesylate (12) can be used to alkylate arylamine (13) to obtain compound A. Alternatively, ketone (6) can be reduced to hydroxy compound (14) using a chiral catalyst such as a Noyori catalyst. The hydroxyl group can be converted to chloride (15) using a chlorinating agent such as 2,4,6-trichloro-1,3,5-triazine. Then, chloride (15) can be used to alkylate arylamine (13) to obtain compound A.

[0060] Definitions As used herein, the terms "treating", "for treating", or "treatment" refer to suppressing, slowing, halting, reducing, shrinking, maintaining a stable medical condition, or reversing the progression or severity of an existing symptom, disorder, condition, or disease.

[0061] As used herein, the term "patient" refers to a mammal, preferably a human.

[0062] As used herein, the terms "cancer" and "cancerous" refer to or describe a physiological condition of a patient typically characterized by uncontrolled cell growth. This definition includes both benign and malignant cancers.

[0063] As used herein, the terms "advanced" or "metastatic" mean cancer that has spread to one or more parts of the body that were not at the site of the original cancerous tissue.

[0064] As used herein, the term "effective amount" refers to the amount or dosage of a therapeutic agent, or a pharmaceutically acceptable salt thereof, such as compound A, or a pharmaceutically acceptable salt thereof, optionally one or more additional agents, or a pharmaceutically acceptable salt thereof, that provides an effective response in a patient being diagnosed or treated.

[0065] As used herein, the terms "effective response" of a patient or "response" of a patient to treatment with a therapeutic agent, or a pharmaceutically acceptable salt thereof, refer to the clinical or therapeutic benefit conferred on the patient when the therapeutic agent, or a pharmaceutically acceptable salt thereof, is administered, optionally in combination with one or more additional agents, or pharmaceutically acceptable salts thereof.

[0066] As used herein, the term "in combination with" means that a therapeutic agent, or a pharmaceutically acceptable salt thereof, and one or more additional therapeutic agents, or pharmaceutically acceptable salts thereof, are administered either separately, simultaneously, or sequentially in any order, for example, at repeated intervals during a standard course of treatment of a single cycle or more than one cycle, such that one agent can be administered before, simultaneously with, or after the administration of the other agent, or administered in any combination thereof.

[0067] As used herein, the term "tromethamine" may otherwise be referred to as tris(hydroxymethyl)aminomethane or tris.

[0068] As used herein, the term "elbamine" may otherwise be referred to as tert-butylamine in other cases.

[0069] Exemplary embodiments Various embodiments of the invention are described in the following numbered clauses.

[0070] Clause 1.2 - A compound that is [[(1R)-1-(3,6-dimethyl-4-oxo-2-phenyl-chromen-8-yl)ethyl]amino]benzoic acid; or a pharmaceutically acceptable salt thereof.

[0071] Clause 2.2 - A compound that is [[(1R)-1-(3,6-dimethyl-4-oxo-2-phenyl-chromen-8-yl)ethyl]amino]benzoic acid.

[0072] Item 3. A compound that is crystalline 2-[[(1R)-1-(3,6-dimethyl-4-oxo-2-phenyl-chromen-8-yl)ethyl]amino]benzoic acid.

[0073] Item 4. A compound according to any one of Items 1 to 3, characterized by an X-ray powder diffraction pattern using CuKα radiation and having at least one peak at diffraction angles 2θ of 7.8° ± 0.2°, 12.1° ± 0.2°, 13.7° ± 0.2°, 14.1° ± 0.2°, 16.8° ± 0.2°, 17.5° ± 0.2°, 18.2° ± 0.2°, 18.9° ± 0.2°, 19.5° ± 0.2°, 20.7° ± 0.2°, 21.2° ± 0.2°, and 24.1° ± 0.2°.

[0074] Item 5. A compound according to any one of Items 1 to 3, characterized by an X-ray powder diffraction pattern using CuKα radiation and having a peak at a diffraction angle 2θ of 12.1° ± 0.2° together with at least one peak selected from 14.1° ± 0.2°, 16.8° ± 0.2°, 18.9° ± 0.2°, and 20.7° ± 0.2°.

[0075] Item 6. A compound according to any one of Items 1 to 3, characterized by an X-ray powder diffraction pattern using CuKα radiation and having a peak at a diffraction angle 2θ of 12.1° ± 0.2° together with at least two peaks selected from 14.1° ± 0.2°, 16.8° ± 0.2°, 18.9° ± 0.2°, and 20.7° ± 0.2°.

[0076] Item 7. A compound according to any one of Items 1 to 3, characterized by an X-ray powder diffraction pattern using CuKα radiation and having a peak at a diffraction angle 2θ of 12.1° ± 0.2° together with at least three peaks selected from 14.1° ± 0.2°, 16.8° ± 0.2°, 18.9° ± 0.2°, and 20.7° ± 0.2°.

[0077] A compound according to any one of clauses 1 to 3, characterized by an X-ray powder diffraction pattern using CuKa radiation, having peaks at diffraction angles 2 theta of 12.1° ± 0.2°, together with peaks of 8.14° ± 0.2°, 16.8° ± 0.2°, 18.9° ± 0.2°, and 20.7° ± 0.2°.

[0078] A compound according to any one of clauses 1 to 3, characterized by an X-ray powder diffraction pattern using CuKa radiation, having peaks at diffraction angles 2 theta of 7.0° ± 0.2°, 9.7° ± 0.2°, 11.9° ± 0.2°, 13.7° ± 0.2°, 14.1° ± 0.2°, 16.8° ± 0.2°, 17.5° ± 0.2°, 18.2° ± 0.2°, 18.9° ± 0.2°, 19.5° ± 0.2°, 20.7° ± 0.2°, 21.2° ± 0.2°, and 24.1° ± 0.2°.

[0079] A compound according to any one of clauses 1 to 3, substantially having the X-ray powder diffraction pattern shown in Figure 1.

[0080] A compound according to any one of clauses 1 to 3, characterized by an X-ray powder diffraction pattern using CuKa radiation, having at least one peak at a diffraction angle 2 theta selected from 7.0° ± 0.2°, 9.7° ± 0.2°, 11.9° ± 0.2°, 14.9° ± 0.2°, and 17.4° ± 0.2°.

[0081] A compound according to any one of clauses 1 to 3, characterized by an X-ray powder diffraction pattern using CuKa radiation, having a peak at a diffraction angle 2 theta of 9.7° ± 0.2°, together with at least one peak selected from 14.9° ± 0.2°, 11.9° ± 0.2°, 17.4° ± 0.2°, and 7.0° ± 0.2°.

[0082] A compound according to any one of clauses 1 to 3, characterized by an X-ray powder diffraction pattern using CuKa radiation, having a peak at a diffraction angle 2 theta of 9.7° ± 0.2°, together with at least two peaks selected from 14.9° ± 0.2°, 11.9° ± 0.2°, 17.4° ± 0.2°, and 7.0° ± 0.2°.

[0083] The compound according to any one of Clauses 1 to 3, characterized by an X-ray powder diffraction pattern using CuKa radiation, having a peak at a diffraction angle 2 theta of 9.7° ± 0.2°, together with at least three peaks selected from 14.14.9° ± 0.2°, 11.9° ± 0.2°, 17.4° ± 0.2°, and 7.0° ± 0.2°.

[0084] The compound according to any one of Clauses 1 to 3, characterized by an X-ray powder diffraction pattern using CuKa radiation, having peaks at diffraction angles 2 theta of 7.0° ± 0.2°, 9.7° ± 0.2°, 11.9° ± 0.2°, 14.9° ± 0.2°, and 17.4° ± 0.2°.

[0085] The compound according to any one of Clauses 1 to 3, substantially having the X-ray powder diffraction pattern shown in Figure 2.

[0086] The compound according to any one of Clauses 1 to 3, characterized by an X-ray powder diffraction pattern using CuKa radiation, having at least one peak at a diffraction angle 2 theta selected from 7.4° ± 0.2°, 8.5° ± 0.2°, 10.6° ± 0.2°, 13.4° ± 0.2°, and 15.7° ± 0.2°.

[0087] The compound according to any one of Clauses 1 to 3, characterized by an X-ray powder diffraction pattern using CuKa radiation, having a peak at a diffraction angle 2 theta of 13.4° ± 0.2°, together with at least one peak selected from 8.5° ± 0.2°, 15.7° ± 0.2°, 10.6° ± 0.2°, and 7.4° ± 0.2°.

[0088] The compound according to any one of Clauses 1 to 3, characterized by an X-ray powder diffraction pattern using CuKa radiation, having a peak at a diffraction angle 2 theta of 13.4° ± 0.2°, together with at least two peaks selected from 8.5° ± 0.2°, 15.7° ± 0.2°, 10.6° ± 0.2°, and 7.4° ± 0.2°.

[0089] The compound according to any one of Clauses 1 to 3, characterized by an X-ray powder diffraction pattern using CuKα radiation, having peaks at diffraction angles 2θ of 13.4° ± 0.2° together with at least three peaks selected from 20.8° ± 0.2°, 15.7° ± 0.2°, 10.6° ± 0.2°, and 7.4° ± 0.2°.

[0090] The compound according to any one of Clauses 1 to 3, characterized by an X-ray powder diffraction pattern using CuKα radiation, having peaks at diffraction angles 2θ of 21.7.4° ± 0.2°, 8.5° ± 0.2°, 10.6° ± 0.2°, 13.4° ± 0.2°, and 15.7° ± 0.2°.

[0091] The compound according to any one of Clauses 1 to 3, substantially having the X-ray powder diffraction pattern shown in Figure 3.

[0092] Clause 23. Tromethamine salt of 2-[[(1R)-1-(3,6-dimethyl-4-oxo-2-phenyl-chromen-8-yl)ethyl]amino]benzoic acid.

[0093] Clause 24. The tromethamine salt according to Clause 23, which is crystalline.

[0094] The tromethamine salt according to Clause 23 or Clause 24, characterized by an X-ray powder diffraction pattern using CuKα radiation, having at least one peak at diffraction angle 2θ selected from 6.4° ± 0.2°, 8.4° ± 0.2°, 10.9° ± 0.2°, 11.8° ± 0.2°, 13.0° ± 0.2°, 16.5° ± 0.2°, 16.9° ± 0.2°, 22.1° ± 0.2°, 23.0° ± 0.2°, and 24.9° ± 0.2°.

[0095] The tromethamine salt according to Clause 23 or Clause 24, characterized by an X-ray powder diffraction pattern using CuKα radiation, having a peak at diffraction angle 2θ of 6.4° ± 0.2° together with at least one peak selected from 8.4° ± 0.2°, 10.9° ± 0.2°, 16.9° ± 0.2°, and 22.1° ± 0.2°.

[0096] The tromethamine salt according to clause 23 or clause 24, characterized by an X-ray powder diffraction pattern using CuKa radiation, having a peak at a diffraction angle 2 theta of 6.4° ± 0.2°, together with at least two peaks selected from 8.4° ± 0.2°, 10.9° ± 0.2°, 16.9° ± 0.2°, and 22.1° ± 0.2°.

[0097] The tromethamine salt according to clause 23 or clause 24, characterized by an X-ray powder diffraction pattern using CuKa radiation, having a peak at a diffraction angle 2 theta of 6.4° ± 0.2°, together with at least three peaks selected from 8.4° ± 0.2°, 10.9° ± 0.2°, 16.9° ± 0.2°, and 22.1° ± 0.2°.

[0098] The tromethamine salt according to clause 23 or clause 24, characterized by an X-ray powder diffraction pattern using CuKa radiation, having a peak at a diffraction angle 2 theta of 6.4° ± 0.2°, together with peaks at 8.4° ± 0.2°, 10.9° ± 0.2°, 16.9° ± 0.2° and 22.1° ± 0.2°.

[0099] The tromethamine salt according to clause 23 or clause 24, characterized by an X-ray powder diffraction pattern using CuKa radiation, having peaks at diffraction angles 2 theta of 6.4° ± 0.2°, 8.4° ± 0.2°, 10.9° ± 0.2°, 11.8° ± 0.2°, 13.0° ± 0.2°, 16.5° ± 0.2°, 16.9° ± 0.2°, 22.1° ± 0.2°, 23.0° ± 0.2° and 24.9° ± 0.2°.

[0100] The tromethamine salt according to clause 23 or clause 24, having substantially the X-ray powder diffraction pattern shown in Figure 4.

[0101] Containing at least one peak based on glycine (external standard at 176.5 ppm) selected from 32.179.0, 158.7, 151.7, 149.7, 136.3, 134.7, 132.9, 129.3, 127.4, 125.2, 121.7, 117.0, 115.5, 115.2, 110.4, 64.1, 63.2, 45.3, 22.6, 20.3, and 11.6 ppm (each ±0.2 ppm). 13 The tromethamine salt according to any one of clauses 23 to 31, characterized by a 13C solid NMR (100.6 MHz) spectrum.

[0102] Containing at least one peak based on glycine (external standard at 176.5 ppm) selected from 33.179.0, 129.3, 63.2, 20.3, and 11.6 ppm (each ±0.2 ppm). 13 The tromethamine salt according to any one of clauses 23 to 31, characterized by a 13C solid NMR (100.6 MHz) spectrum.

[0103] Clause 34. Containing peaks based on glycine (external standard at 176.5 ppm) at 179.0, 129.3, 63.2, 20.3, and 11.6 ppm (each ±0.2 ppm). C The tromethamine salt according to any one of clauses 23 to 31, characterized by a 13C solid NMR (100.6 MHz) spectrum.

[0104] Clause 35. Containing peaks based on glycine (external standard at 176.5 ppm) at 179.0, 158.7, 151.7, 149.7, 136.3, 134.7, 132.9, 129.3, 127.4, 125.2, 121.7, 117.0, 115.5, 115.2, 110.4, 64.1, 63.2, 45.3, 22.6, 20.3, and 11.6 ppm (each ±0.2 ppm). 13 The tromethamine salt according to any one of clauses 23 to 31, characterized by a 13C solid NMR (100.6 MHz) spectrum.

[0105] The tromethamine salt according to clause 23 or clause 24, characterized by an X-ray powder diffraction pattern using CuKa radiation, having at least one peak at diffraction angles 2 theta selected from 10.6° ± 0.2°, 13.2° ± 0.2°, 14.5° ± 0.2°, 15.9° ± 0.2°, and 17.4° ± 0.2°.

[0106] The tromethamine salt according to clause 23 or clause 24, characterized by an X-ray powder diffraction pattern using CuKa radiation, having a peak at a diffraction angle 2 theta of 15.9° ± 0.2° together with at least one peak selected from 10.6° ± 0.2°, 17.4° ± 0.2°, 13.2° ± 0.2°, and 14.5° ± 0.2°.

[0107] The tromethamine salt according to clause 23 or clause 24, characterized by an X-ray powder diffraction pattern using CuKa radiation, having a peak at a diffraction angle 2 theta of 15.9° ± 0.2° together with at least two peaks selected from 10.6° ± 0.2°, 17.4° ± 0.2°, 13.2° ± 0.2°, and 14.5° ± 0.2°.

[0108] The tromethamine salt according to clause 23 or clause 24, characterized by an X-ray powder diffraction pattern using CuKa radiation, having a peak at a diffraction angle 2 theta of 15.9° ± 0.2° together with at least three peaks selected from 10.6° ± 0.2°, 17.4° ± 0.2°, 13.2° ± 0.2°, and 14.5° ± 0.2°.

[0109] The tromethamine salt according to clause 23 or clause 24, characterized by an X-ray powder diffraction pattern using CuKa radiation, having peaks at diffraction angles 2 theta of 10.6° ± 0.2°, 13.2° ± 0.2°, 14.5° ± 0.2°, 15.9° ± 0.2° and 17.4° ± 0.2°.

[0110] The tromethamine salt according to clause 23 or clause 24, substantially having the X-ray powder diffraction pattern shown in Figure 5.

[0111] The tromethamine salt according to clause 23 or clause 24, characterized by an X-ray powder diffraction pattern using CuKa radiation and having at least one peak at diffraction angles 2 theta selected from 42.6° ± 0.2°, 11.1° ± 0.2°, 12.6° ± 0.2°, 17.1° ± 0.2°, and 18.9° ± 0.2°.

[0112] The tromethamine salt according to clause 23 or clause 24, characterized by an X-ray powder diffraction pattern using CuKa radiation and having a peak at a diffraction angle 2 theta of 11.1° ± 0.2° together with at least one peak selected from 12.6° ± 0.2°, 17.1° ± 0.2°, 6.3° ± 0.2°, and 18.9° ± 0.2°.

[0113] The tromethamine salt according to clause 23 or clause 24, characterized by an X-ray powder diffraction pattern using CuKa radiation and having a peak at a diffraction angle 2 theta of 11.1° ± 0.2° together with at least two peaks selected from 12.6° ± 0.2°, 17.1° ± 0.2°, 6.3° ± 0.2°, and 18.9° ± 0.2°.

[0114] The tromethamine salt according to clause 23 or clause 24, characterized by an X-ray powder diffraction pattern using CuKa radiation and having a peak at a diffraction angle 2 theta of 11.1° ± 0.2° together with at least three peaks selected from 12.6° ± 0.2°, 17.1° ± 0.2°, 6.3° ± 0.2°, and 18.9° ± 0.2°.

[0115] The tromethamine salt according to clause 23 or clause 24, characterized by an X-ray powder diffraction pattern using CuKa radiation and having peaks at diffraction angles 2 theta of 6.3° ± 0.2°, 11.1° ± 0.2°, 12.6° ± 0.2°, 17.1° ± 0.2° and 18.9° ± 0.2°.

[0116] The tromethamine salt according to clause 23 or clause 24, substantially having the X-ray powder diffraction pattern shown in Figure 6.

[0117] Item 48.2 - The albumin salt of [[(1R)-1-(3,6-dimethyl-4-oxo-2-phenyl-chromen-8-yl)ethyl]amino]benzoic acid.

[0118] Item 49. The albumin salt according to Item 48, which is crystalline.

[0119] Item 50. The albumin salt according to Item 48 or Item 49, characterized by an X-ray powder diffraction pattern using CuKa radiation and having at least one peak at diffraction angle 2 theta selected from 6.5° ± 0.2°, 10.5° ± 0.2°, 11.1° ± 0.2°, 15.2° ± 0.2°, 15.9° ± 0.2°, 17.6° ± 0.2°, 18.0° ± 0.2°, 19.3° ± 0.2°, 21.5° ± 0.2°, 22.2° ± 0.2°, 22.7° ± 0.2°, and 26.3° ± 0.2°.

[0120] Item 51. The albumin salt according to Item 48 or Item 49, characterized by an X-ray powder diffraction pattern using CuKa radiation and having a peak at diffraction angle 2 theta of 11.1° ± 0.2° together with at least one peak selected from 10.5° ± 0.2°, 15.2° ± 0.2°, 18.0° ± 0.2°, and 19.3° ± 0.2°.

[0121] Item 52. The albumin salt according to Item 48 or Item 49, characterized by an X-ray powder diffraction pattern using CuKa radiation and having a peak at diffraction angle 2 theta of 11.1° ± 0.2° together with at least two peaks selected from 10.5° ± 0.2°, 15.2° ± 0.2°, 18.0° ± 0.2°, and 19.3° ± 0.2°.

[0122] Item 53. The albumin salt according to Item 48 or 49, characterized by an X-ray powder diffraction pattern using CuKa radiation and having a peak at diffraction angle 2 theta of 11.1° ± 0.2° together with at least three peaks selected from 10.5° ± 0.2°, 15.2° ± 0.2°, 18.0° ± 0.2°, and 19.3° ± 0.2°.

[0123] The elbumin salt according to clause 48 or clause 49, characterized by an X-ray powder diffraction pattern using CuKa radiation, having peaks at diffraction angles 2 theta of 11.1° ± 0.2°, along with peaks of 54.10.5° ± 0.2°, 15.2° ± 0.2°, 18.0° ± 0.2° and 19.3° ± 0.2°.

[0124] The elbumin salt according to clause 48 or clause 49, characterized by an X-ray powder diffraction pattern using CuKa radiation, having peaks at diffraction angles 2 theta of 66.5° ± 0.2°, 10.5° ± 0.2°, 11.1° ± 0.2°, 15.2° ± 0.2°, 15.9° ± 0.2°, 17.6° ± 0.2°, 18.0° ± 0.2°, 19.3° ± 0.2°, 21.5° ± 0.2°, 22.2° ± 0.2°, 22.7° ± 0.2°, and 26.3° ± 0.2°.

[0125] The elbumin salt according to clause 48 or clause 49, substantially having the X-ray powder diffraction pattern shown in Figure 7.

[0126] The elbumin salt according to any one of clauses 48 to 56, characterized by a 13C solid NMR (100.6 MHz) spectrum including at least one peak referenced to glycine (external reference at 176.5 ppm) selected from 177.4, 174.8, 159.8, 151.7, 149.5, 134.0, 132.6, 130.7, 130.3, 129.0, 128.1, 123.1, 122.4, 119.4, 116.8, 115.9, 112.3, 53.0, 47.5, 27.4, 23.3, 21.3, and 11.3 ppm (each ± 0.2 ppm). 13 The elbumin salt according to any one of clauses 48 to 56, characterized by a 13C solid NMR (100.6 MHz) spectrum including at least one peak referenced to glycine (external reference at 176.5 ppm) selected from 177.4, 132.6, 27.4, 21.3, and 11.3 ppm (each ± 0.2 ppm).

[0127] The elbumin salt according to any one of clauses 48 to 56, characterized by a 13C solid NMR (100.6 MHz) spectrum including at least one peak referenced to glycine (external reference at 176.5 ppm) selected from 177.4, 132.6, 27.4, 21.3, and 11.3 ppm (each ± 0.2 ppm). 13 The elbumin salt according to any one of clauses 48 to 56, characterized by a 13C solid NMR (100.6 MHz) spectrum including at least one peak referenced to glycine (external reference at 176.5 ppm) selected from 177.4, 132.6, 27.4, 21.3, and 11.3 ppm (each ± 0.2 ppm).

[0128] Containing peaks referenced to glycine (external standard at 176.5 ppm) at 59.177.4, 132.6, 27.4, 21.3, and 11.3 ppm (±0.2 ppm each). 13 An albumin salt according to any one of clauses 48 to 56, characterized by a 13C solid NMR (100.6 MHz) spectrum.

[0129] Containing peaks referenced to glycine (external standard at 176.5 ppm) at 60.177.4, 174.8, 159.8, 151.7, 149.5, 134.0, 132.6, 130.7, 130.3, 129.0, 128.1, 123.1, 122.4, 119.4, 116.8, 115.9, 112.3, 53.0, 47.5, 27.4, 23.3, 21.3, and 11.3 ppm (±0.2 ppm each). 13 An albumin salt according to any one of clauses 48 to 56, characterized by a 13C solid NMR (100.6 MHz) spectrum.

[0130] Clause 61. A pharmaceutical composition comprising a compound according to any one of clauses 1 to 22, a tromethamine salt according to any one of clauses 23 to 47, or an albumin salt according to any one of clauses 48 to 60, and a pharmaceutically acceptable carrier.

[0131] Clause 62. A method of inhibiting phosphoinositide 3-kinase (PI3K), comprising administering to a patient in need thereof a therapeutically effective amount of a compound according to any one of clauses 1 to 22, a tromethamine salt according to any one of clauses 23 to 47, an albumin salt according to any one of clauses 48 to 60, or a pharmaceutical composition according to clause 61.

[0132] Clause 63. A method of treating a patient having a disease associated with mutant phosphoinositide 3-kinase (PI3K), comprising administering to the patient a therapeutically effective amount of a compound according to any one of clauses 1 to 22, a tromethamine salt according to any one of clauses 23 to 47, an albumin salt according to any one of clauses 48 to 60, or a pharmaceutical composition according to clause 61.

[0133] Item 64. The method according to item 62 or item 63, wherein the PI3K is PI3Kα.

[0134] Item 65. The method according to any one of items 62 to 64, wherein the PI3K has the H1047R mutation.

[0135] Item 66. The method according to any one of items 63 to 65, wherein the disease is cancer.

[0136] Item 67. The method according to item 66, wherein the cancer is endometrial cancer, gastric cancer, leukemia, lymphoma, sarcoma, colorectal cancer, lung cancer, ovarian cancer, skin cancer, head and neck cancer, breast cancer, brain cancer, or prostate cancer.

[0137] Item 68. The method according to item 66, wherein the cancer is breast cancer.

[0138] Item 69. The method according to item 66, wherein the cancer is hormone receptor positive (HR+), human epidermal growth factor receptor 2 negative (HER2−) advanced or metastatic breast cancer.

[0139] Item 70. The method according to any one of items 63 to 65, wherein the disease is CLOVES syndrome (congenital lipomatous overgrowth, vascular malformation, epidermal nevus, scoliosis / skeletal and spinal syndrome), or PIK3CA-related overgrowth syndrome (PROS).

[0140] Item 71. A method for treating a patient having a PIK3CA mutant cancer, comprising administering to the patient an effective amount of the compound according to any one of items 1 to 22, the tromethamine salt according to any one of items 23 to 47, the albumin salt according to any one of items 48 to 60, or the pharmaceutical composition according to item 61.

[0141] Article 72. A method for treating a patient having a PIK3CA - mutated solid tumor, comprising administering to the patient an effective amount of the compound according to any one of Articles 1 to 22, the tromethamine salt according to any one of Articles 23 to 47, the albumin salt according to any one of Articles 48 to 60, or the pharmaceutical composition according to Article 61. A method for treating a patient having a PIK3CA - mutated solid tumor.

[0142] Article 73. A method for treating a patient having a PIK3CA - mutated breast cancer, comprising administering to the patient an effective amount of the compound according to any one of Articles 1 to 22, the tromethamine salt according to any one of Articles 23 to 47, the albumin salt according to any one of Articles 48 to 60, or the pharmaceutical composition according to Article 61. A method for treating a patient having a PIK3CA - mutated breast cancer.

[0143] Article 74. A method for treating a patient having a PIK3CA - mutated, progressive or metastatic breast cancer, comprising administering to the patient an effective amount of the compound according to any one of Articles 1 to 22, the tromethamine salt according to any one of Articles 23 to 47, the albumin salt according to any one of Articles 48 to 60, or the pharmaceutical composition according to Article 61. A method for treating a patient having a PIK3CA - mutated, progressive or metastatic breast cancer.

[0144] Article 75. The method according to Article 71, wherein the PIK3CA - mutated cancer is a PIK3CA H1047R - mutated cancer.

[0145] Article 76. The method according to Article 72, wherein the PIK3CA - mutated solid tumor is a PIK3CA H1047R - mutated solid tumor.

[0146] Article 77. The method according to Article 72, wherein the PIK3CA - mutated solid tumor is selected from gynecological cancer, head and neck cancer, and triple - negative breast cancer.

[0147] Article 78. The method according to Article 77, wherein the PIK3CA - mutated solid tumor is a gynecological cancer.

[0148] Article 79. The method according to Article 77, wherein the solid tumor with PIK3CA mutation is a head and neck cancer.

[0149] Article 80. The method according to Article 77, wherein the solid tumor with PIK3CA mutation is a triple-negative breast cancer.

[0150] Article 81. The method according to Article 73, wherein the breast cancer with PIK3CA mutation is a breast cancer with PIK3CA H1047R mutation.

[0151] Article 82. The method according to Article 74, wherein the breast cancer with PIK3CA mutation, progressive or metastatic, is a breast cancer with PIK3CA H1047R mutation, progressive or metastatic.

[0152] Article 83. The method according to Article 74, wherein the breast cancer with PIK3CA mutation, progressive or metastatic, is estrogen receptor positive (ER+), human epidermal growth factor receptor 2 negative (HER2-), and has a PIK3CA mutation, progressive or metastatic.

[0153] Article 84. The method according to Article 74, wherein the breast cancer with PIK3CA mutation, progressive or metastatic, is estrogen receptor positive (ER+), human epidermal growth factor receptor 2 negative (HER2-), and has a PIK3CA H1047R mutation, progressive or metastatic.

[0154] Article 85. A compound according to any one of Articles 1 to 22, a tromethamine salt according to any one of Articles 23 to 47, an albumin salt according to any one of Articles 48 to 60, or a pharmaceutical composition according to Article 61, for use in treatment.

[0155] Article 86. A compound according to any one of Articles 1 to 22, a tromethamine salt according to any one of Articles 23 to 47, an albumin salt according to any one of Articles 48 to 60, or a pharmaceutical composition according to Article 61, for use in the treatment of diseases associated with mutant phosphoinositide 3-kinase (PI3K).

[0156] A compound, tromethamine salt, albumin salt, or pharmaceutical composition for use according to item 86, wherein the PI3K is PI3Kα.

[0157] A compound, tromethamine salt, albumin salt, or pharmaceutical composition for use according to item 86 or 87, wherein the PI3K has the H1047R mutation.

[0158] A compound, tromethamine salt, albumin salt, or pharmaceutical composition for use according to any one of items 86 to 88, wherein the disease is cancer.

[0159] A compound, tromethamine salt, albumin salt, or pharmaceutical composition for use according to item 89, wherein the cancer is endometrial cancer, gastric cancer, leukemia, lymphoma, sarcoma, colorectal cancer, lung cancer, ovarian cancer, skin cancer, head and neck cancer, breast cancer, brain cancer, or prostate cancer.

[0160] A compound, tromethamine salt, albumin salt, or pharmaceutical composition for use according to item 89, wherein the cancer is breast cancer.

[0161] A compound, tromethamine salt, albumin salt, or pharmaceutical composition for use according to item 89, wherein the cancer is hormone receptor positive (HR+), human epidermal growth factor receptor 2 negative (HER2-) advanced or metastatic breast cancer.

[0162] A compound, tromethamine salt, albumin salt, or pharmaceutical composition for use according to any one of items 86 to 88, wherein the disease is CLOVES syndrome (congenital lipomatous overgrowth, vascular malformations, epidermal nevi, scoliosis / skeletal and spinal syndrome), or PIK3CA-related overgrowth syndrome (PROS).

[0163] A compound according to any one of items 1 to 22, a tromethamine salt according to any one of items 23 to 47, an albumin salt according to any one of items 48 to 60, or a pharmaceutical composition according to item 61 for use in the treatment of PIK3CA mutant cancer.

[0164] Article 95. A compound according to any one of Articles 1 to 22, a tromethamine salt according to any one of Articles 23 to 47, an albumin salt according to any one of Articles 48 to 60, or a pharmaceutical composition according to Article 61, for use in the treatment of PIK3CA mutant solid tumors.

[0165] Article 96. A compound according to any one of Articles 1 to 22, a tromethamine salt according to any one of Articles 23 to 47, an albumin salt according to any one of Articles 48 to 60, or a pharmaceutical composition according to Article 61, for use in the treatment of PIK3CA mutant breast cancer.

[0166] Article 97. A compound according to any one of Articles 1 to 22, a tromethamine salt according to any one of Articles 23 to 47, an albumin salt according to any one of Articles 48 to 60, or a pharmaceutical composition according to Article 61, for use in the treatment of PIK3CA mutant, advanced or metastatic breast cancer.

[0167] Article 98. A compound, tromethamine salt, albumin salt, or pharmaceutical composition according to Article 94, for use in the treatment according to Article 94, wherein the PIK3CA mutant cancer is a PIK3CA H1047R mutant cancer.

[0168] Article 99. A compound, tromethamine salt, albumin salt, or pharmaceutical composition according to Article 95, for use in the treatment according to Article 95, wherein the PIK3CA mutant solid tumor is a PIK3CA H1047R mutant solid tumor.

[0169] Article 100. A compound, tromethamine salt, albumin salt, or pharmaceutical composition according to Article 95, for use in the treatment according to Article 95, wherein the PIK3CA mutant solid tumor is selected from gynecological cancer, head and neck cancer, and triple-negative breast cancer.

[0170] Article 101. A compound, tromethamine salt, albumin salt, or pharmaceutical composition according to Article 100, for use in the treatment according to Article 100, wherein the PIK3CA mutant solid tumor is a gynecological cancer.

[0171] The compound, tromethamine salt, albumin salt, or pharmaceutical composition for the use according to clause 100, wherein the solid tumor with PIK3CA mutation is a head and neck cancer.

[0172] The compound, tromethamine salt, albumin salt, or pharmaceutical composition for the use according to clause 100, wherein the solid tumor with PIK3CA mutation is a triple-negative breast cancer.

[0173] The compound, tromethamine salt, albumin salt, or pharmaceutical composition for the use according to clause 96, wherein the breast cancer with PIK3CA mutation is a breast cancer with PIK3CA H1047R mutation.

[0174] The compound, tromethamine salt, albumin salt, or pharmaceutical composition for the use according to clause 97, wherein the breast cancer with PIK3CA mutation, progressive or metastatic breast cancer is a progressive or metastatic breast cancer with PIK3CA H1047R mutation.

[0175] The compound, tromethamine salt, albumin salt, or pharmaceutical composition for the use according to clause 97, wherein the breast cancer with PIK3CA mutation, progressive or metastatic breast cancer is an estrogen receptor-positive (ER+), human epidermal growth factor receptor 2-negative (HER2-), breast cancer with PIK3CA mutation, progressive or metastatic breast cancer.

[0176] The compound, tromethamine salt, albumin salt, or pharmaceutical composition for the use according to clause 97, wherein the breast cancer with PIK3CA mutation, progressive or metastatic breast cancer is an estrogen receptor-positive (ER+), human epidermal growth factor receptor 2-negative (HER2-), breast cancer with PIK3CA H1047R mutation, progressive or metastatic breast cancer.

[0177] Use of the compound according to any one of clauses 1 to 22, the tromethamine salt according to any one of clauses 23 to 47, the albumin salt according to any one of clauses 48 to 60, or the pharmaceutical composition according to clause 61, in the manufacture of a medicament for treating breast cancer with PIK3CA mutation.

[0178] Use of a compound as described in any one of Clauses 1 to 22, a tromethamine salt as described in any one of Clauses 23 to 47, an albumin salt as described in any one of Clauses 48 to 60, or a pharmaceutical composition as described in Clause 61, in the manufacture of a medicament for treating PIK3CA-mutated solid tumors.

[0179] Use of a compound as described in any one of Clauses 1 to 22, a tromethamine salt as described in any one of Clauses 23 to 47, an albumin salt as described in any one of Clauses 48 to 60, or a pharmaceutical composition as described in Clause 61, in the manufacture of a medicament for treating PIK3CA-mutated breast cancer.

[0180] Use of a compound as described in any one of Clauses 1 to 22, a tromethamine salt as described in any one of Clauses 23 to 47, an albumin salt as described in any one of Clauses 48 to 60, or a pharmaceutical composition as described in Clause 61, in the manufacture of a medicament for treating PIK3CA-mutated, advanced or metastatic breast cancer.

Examples

[0181] Nuclear magnetic resonance (NMR) spectra were recorded at 400 MHz or 300 MHz and 300.3 K as described, unless otherwise specified. Chemical shifts (δ) are reported in parts per million (ppm). Spectra were recorded using a Bruker or Varian instrument with 8, 16, or 32 scans.

[0182] LC-MS chromatograms and spectra were recorded using an Agilent 1200 or Shimadzu LC-20 AD & MS 2020 instrument with a C-18 column such as Luna-C18 2.0×30 mm or Xbridge Shield RPC18 2.1×50 mm. The injection volume was 0.7 - 8.0 μl and the flow rate was typically 0.8 or 1.2 ml / min. The positive detection methods were diode array (DAD) or evaporative light scattering (ELSD) and positive electrospray ionization. The MS range was 100 - 1000 Da. The solvents were all gradients of water and acetonitrile containing a modifier such as trifluoroacetic acid or ammonium carbonate (typically 0.01 - 0.04%).

[0183] The XRPD pattern of the crystalline solid was obtained on a Bruker D8 Endeavor X-ray powder diffractometer operating at 40 kV and 40 mA, equipped with a CuKα (1.5418 Å) source and a Linxeye detector. The sample was scanned at 4–42° 2θ with a step size of 0.009 2θ° and a scan speed of 0.5 s / step, using a 0.3° primary slit aperture and a 3.9° particle size distribution (PSD) aperture, or scanned at 4–30 2θ° with a step size of 0.009 2θ° and a scan speed of 0.25 s / step, using a 0.3° primary slit aperture and a 3.9° particle size distribution (PSD) aperture. The dry powder was filled into a quartz sample holder, and a smooth surface was obtained using a glass slide. The crystalline form diffraction pattern was collected at ambient temperature and relative humidity. The crystalline peak positions were determined with MDI-Jade after global pattern shifting based on an internal NIST 675 standard having peaks at 8.853 and 26.774 2θ°. In the field of crystallography, it is well known that for any given crystalline form, the relative intensities of diffraction peaks can vary due to preferred orientations arising from factors such as crystalline form and crystal habit. When the effect of preferred orientation is present, the peak intensities are modified, but the characteristic peak positions of the polymorphs remain invariant. See, for example, The United States Pharmacopeia#23, National Formulary#18, pages 1843–1844, 1995. Further, it is also well known in the field of crystallography that for any given crystalline form, the angular peak positions can vary slightly. For example, the peak positions can be shifted by temperature fluctuations at which the sample is analyzed, sample displacement, or the presence or absence of an internal standard. In this case, a peak position variability of ±0.2 2θ° is estimated to account for these potential variations without preventing a clear identification of the indicated crystalline form. Confirmation of the crystalline form can be made based on any unique combination of characteristic peaks.

[0184] Preliminary inspection and data collection of single crystal X-ray diffraction were performed on a Rigaku SuperNova diffractometer equipped with a copper anode microfocus sealed X-ray tube (Cu Kα λ = 1.54184 Å) and a Dectris Pilatus3 R 200K hybrid pixel array detector. Cell refinement and data reduction were achieved using CRYSALISPRO (CrysAlisPro 1.171.38.41r (Rigaku Oxford Diffraction, 2015)). Data were collected at room temperature up to a maximum diffraction angle (2θ) of 151.16° or 151.992°. The structure was solved by the direct method using SHELXT (Sheldrick, G. M. Acta Cryst. 2015, A71, 3 - 8). The structure was refined using SHELXL - 2014 (Sheldrick, G. M. Acta Cryst. 2008, A64, 112 - 122). Hydrogen atoms present on nitrogen were refined independently. All other hydrogen atoms were included in the refinement but were constrained to ride on the atoms to which they were bonded. The structure was refined by full-matrix least squares. The simulated XRPD pattern was generated using unit cell parameters and atomic coordinates from the single crystal structure in MERCURY crystal modeling software.

[0185] Differential scanning calorimetry (DSC) analysis was performed using a TA Q2000 DSC executed by TA Thermal Advantage Software v5.2.6, and the data were analyzed by Universal Analysis 2000 v4.5a. Samples were equilibrated at 25 °C in crimped aluminum pans and then pierced prior to heating at 10 °C / min to 300 °C using a nitrogen purge of 50 mL / min. Temperature and heat flow were calibrated against indium melting.

[0186] Thermogravimetric analysis was performed using a TA Instruments Q5000 TGA run by TA Thermal Advantage Software v5.2.6, and the data was analyzed by Universal Analysis 2000 v4.5a. Samples (3 - 10 mg) were heated from ambient temperature (ca. 25 °C) to 200 °C at a rate of 10 °C / min. N2 was used as the carrier (10 mL / min) and purge (50 mL / min) gas. The temperature was calibrated by Curie temperature measurement using nickel and alumel standards. Weight calibration was performed using standards supplied by the manufacturer.

[0187] Solid-state NMR (ssNMR) spectra were obtained on an Agilent DD2-400 NMR spectrometer and processed with VnmrJ v3.2A. The data was externally referenced to glycine at 176.5 ppm.

[0188] ssNMR parameters

Table 1

[0189] Abbreviations: ACN Acetonitrile AcOH Acetic acid ADP Adenosine diphosphate ATP Adenosine triphosphate CDCl3 Chloroform-d DCM Dichloromethane DMF N,N-Dimethylformamide DMSO Dimethyl sulfoxide DMSO-d6 Hexadeuterodimethyl sulfoxide DSC Differential scanning calorimetry eq Equivalent EtOAc Ethyl acetate EtOH Ethanol h Hour(s) 1 1H NMR Proton nuclear magnetic resonance spectroscopy IPA Isopropanol Kg Kilogram L liter LC-MS Liquid Chromatography-Mass Spectrometry MeOH Methanol MPa Megapascal 2-MeTHF 2-Methyltetrahydrofuran min minute(s) MS ES Mass Spectrometry Electrospray ppm Parts per million rt Room temperature SFC Supercritical Fluid Chromatography THF Tetrahydrofuran TGA Thermogravimetric Analysis XRD X-Ray Diffraction XRPD Powder X-Ray Diffraction

[0190] Example 1 2-[[(1R)-1-(3,6-Dimethyl-4-oxo-2-phenyl-chromen-8-yl)ethyl]amino]benzoic acid (「Compound A」) Intermediate 1: (2-Bromo-4-methyl-phenyl)propanoate

[0191]

Chemical formula

[0192] Intermediate 2: 1-(3-Bromo-2-hydroxy-5-methyl-phenyl)propan-1-one

[0193]

Chem.

[0194] Route 2: (2-Bromo-4-methyl-phenyl)propanoate (120 g, 496 mmol) was transferred to a reactor, cooled to -20 °C, and treated with trifluoromethanesulfonic acid (216 mL). After the addition was complete, the reaction was stirred at 60 °C for 1 h. The reaction was cooled to room temperature and poured into ice water (600 mL). The product was removed as a yellow solid by filtration (114 g, 95%). MS ES - m / z 241, 243 [M - H] - . 1 H NMR (400 MHz, CDCl3) δ ppm 1.26 (t, 3H), 2.33 (s, 3H), 3.06 (q, 2H), 7.55 (m, 2H), 12.87 (s, 1H).

[0195] Intermediate 3: (E)-1-(3-Bromo-2-hydroxy-5-methyl-phenyl)-2-methyl-3-phenyl-prop-2-en-1-one

[0196]

Chem.

[0197] Intermediate 4: 8-Bromo-3,6-dimethyl-2-phenyl-chromen-4-one

[0198]

Chem.

[0199] Route 2: A solution of 1-(3-bromo-2-hydroxy-5-methyl-phenyl)propan-1-one (50.0 g, 205.7 mmol) in THF (100 mL) was cooled to -80 °C and treated with lithium bis(trimethylsilyl)amide (1 M in THF, 617 mmol). After stirring at -80 °C for 1 hour, the mixture was warmed to 0 °C and stirred for 1 hour. The mixture was cooled to -80 °C and treated dropwise with benzoyl chloride (37.6 g, 267.4 mmol). After the addition was complete, the reaction was stirred at 25 °C for 16 hours. The reaction was cooled to -20 °C and the pH was adjusted to 4 with 50% aqueous acetic acid. The THF was removed under vacuum and the precipitate was removed by filtration. The solid was dissolved in acetic acid and aqueous HCl (250 mL / 10 mL) and the resulting solution was stirred at 100 °C for 1 hour. The mixture was cooled to 20 °C and diluted with water (100 mL). The solid was removed by filtration, washed with water (50 mL) and triturated with 200 mL of EtOAc at room temperature for 30 minutes. The product was recovered as an off-white solid by filtration (170.7 g, 83%). MS ES+ m / z 329,331 [M+H] + . 1 H NMR (400 MHz, CDCl3) δ ppm 2.24 (s, 3H), 2.48 (s, 3H), 7.55 - 7.57 (m, 3H), 7.75 - 7.78 (m, 3H), 8.00 (s, 1H).

[0200] Intermediate 5: 8-Acetyl-3,6-dimethyl-2-phenyl-chromen-4-one

[0201]

Chemical Structure

[0202] Route 2: A mixture of 8-bromo-3,6-dimethyl-2-phenyl-chromen-4-one (50.0 g, 151.9 mmol), palladium acetate (0.34 g, 1.52 mmol), 1,3-bis(diphenylphosphino)propane (1.25 g, 3.04 mmol), triethylamine (46.17 g, 455.7 mmol), n-butyl vinyl ether (76 g, 759.5 mmol), and ethylene glycol (400 mL) was stirred at 100 °C for 7 h under a nitrogen atmosphere. The reaction mixture was cooled to 30 °C, treated with 3 g of activated carbon, and stirred. The suspension was filtered through celite, and the pH of the filtrate was adjusted to 3 - 4 with HCl and stirred at 60 °C for 12 h. The reaction mixture was cooled to 40 °C, and the solid was removed by filtration. The solid was slurried in 250 mL of THF and stirred at 60 °C for 5 h. The reaction mixture was cooled to 30 °C, and the product (40 g, 90%) was removed by filtration and dried at 50 °C to give an off-white solid. MS ES+ m / z 293 [M+H] +. 1H NMR (400 mHz, CDCl3) δ ppm 2.21 (s, 3H), 2.52 (s, 3H), 2.73 (s, 3H), 7.55 - 7.58 (m, 3H), 7.67 - 7.70 (m, 2H), 7.97 (s, 1H), 8.26 - 8.27 (m, 1H).

[0203] Intermediate 6: (NE,R)-N-[1-(3,6-dimethyl-4-oxo-2-phenyl-chromen-8-yl)ethylidene]-2-methyl-propan-2-sulfinamide

[0204]

Chem.

[0205] Intermediate 7: (R)-N-[(1R)-1-(3,6-dimethyl-4-oxo-2-phenyl-chromen-8-yl)ethyl]-2-methyl-propan-2-sulfinamide

[0206]

Chem.

[0207] Intermediate 8: 8-[(1R)-1-Aminoethyl]-3,6-dimethyl-2-phenyl-chromen-4-one

[0208]

Chemical formula

[0209] Intermediate 9: 8-[(1S)-1-Hydroxyethyl]-3,6-dimethyl-2-phenyl-chromen-4-one

[0210]

Chem.

[0211] Intermediate 10: [(1S)-1-(3,6-Dimethyl-4-oxo-2-phenyl-chromen-8-yl)ethyl]methanesulfonate

[0212]

Chem.

[0213] Intermediate 11: 8-[(1R)-1-hydroxyethyl]-3,6-dimethyl-2-phenyl-chromen-4-one

[0214] [Chemical formula] Into a flask equipped with overhead stirring, a condenser and a temperature probe, 8-acetyl-3,6-dimethyl-2-phenyl-chromen-4-one (10 g, 34.2 mmol) and RuCl(p-cymene)[(R,R)-TsDPEN] (CAS 192139-92-7, 0.65 g, 1.03 mmol) were charged. 50 mL of methanol was added and stirring was started. The reaction was cooled to 10 °C and slowly treated with 1,8-diazabicyclo[5.4.0]undec-7-ene (15.62 g, 102.62 mmol) while maintaining the temperature below 25 °C. After the addition was complete, the reaction was cooled back to 10 °C and the reaction was divided and treated with formic acid (4.72 g, 102.62 mmol) while maintaining the temperature below 15 °C. After the addition, the reaction was stirred at 55 °C for about 3 hours. The reaction was cooled to 20 °C, treated with 4M aqueous HCl solution (50 mL) for 1 hour, and the resulting slurry was stirred at room temperature overnight. The product (9.35 g, 95%) was isolated by filtration, washed with water, and dried under vacuum at 45 °C.

[0215] Alternative synthesis of Intermediate 11: 8-[(1R)-1-hydroxyethyl]-3,6-dimethyl-2-phenyl-chromen-4-one THF (3 L / kg, 360 L), 8-acetyl-3,6-dimethyl-2-phenyl-chromen-4-one (120 kg, 410 mol), and sodium methoxide (2.2 kg, 41 mol, 0.10 eq) were charged into the reactor. The reactor was evacuated and refilled with nitrogen three times. (S)-RUCY-XylBINAP [CAS#1312713-89-5] (364 g, 0.31 mol, 0.00075 eq) was charged into the reactor, and the reactor was washed with THF (2 L / kg, 240 L). It was evacuated and refilled with nitrogen three times. The reactor was pressurized with 1 MPa of hydrogen, and the reaction mixture was stirred at 40 °C for 18 h. After cooling to 25 °C, a sample for analysis was taken. If the reaction was not complete, the reactor was refilled with 1 MPa of hydrogen and stirred at 40 °C for an additional 6 h.

[0216] When the reaction was complete, 2-MeTHF (5 L / kg, 600 L) and purified water (5 L / kg, 600 L) were charged into the reactor and stirred at 25 °C for 30 min. The reaction mixture was filtered, and the solid was washed with 2-MeTHF (1 L / kg, 120 L). It was left standing for 30 min. The aqueous layer was removed, and the organic layer was solvent-exchanged into ACN (4 L / kg, 480 L). It was cooled to 5 °C over 10 h, and the mixture was stirred at 5 °C for 6 h. The mixture was filtered, and the solid was washed with ACN (1.52 L / kg, 182 L) and n-heptane (5.0 L / kg, 600 L) and dried under vacuum at 45 °C for 12 h to obtain the title compound (102.7 kg, 85%). 1 H NMR (500 MHz, DMSO-d6) δ ppm 1.40 (d, 3H, J = 6.5 Hz), 2.03 (s, 3H), 2.41 (s, 3H), 5.24 (dq, 1H, J = 4.2, 6.5 Hz), 5.38 (d, 1H, J = 4.2 Hz), 7.58 (m, 3H), 7.73 (m, 4H).

[0217] Intermediate 12: 8-[(1S)-1-chloroethyl]-3,6-dimethyl-2-phenyl-chromen-4-one

[0218]

Chemical Structure

[0219] Alternative synthesis of Intermediate 12: 8-[(1S)-1-chloroethyl]-3,6-dimethyl-2-phenyl-chromen-4-one The reactor was charged with 2-MeTHF (5 L / kg, 480 L) and 8-[(1R)-1-hydroxyethyl]-3,6-dimethyl-2-phenyl-chromen-4-one (96 kg, 326.5 mol) and cooled to 10 °C. The reactor was charged with 1-formylpyrrolidine (32.4 kg, 326.5 mol, 1 equiv), and then benzoyl chloride (91.8 kg, 653 mol, 2 equiv) was added over 2 h. The reaction mixture was warmed to 25 °C and stirred for 18 h. After completion of the reaction, the reaction mixture was diluted with 2-MeTHF (5 L / kg, 480 L) and cooled to 15 °C. 3M aqueous NaOH solution (5 L / kg, 480 L) was added slowly, the mixture was warmed to 25 °C, and the mixture was stirred for 45 min. After standing for 30 min, the aqueous layer was removed. Water (5 L / kg, 480 L) was charged to the reactor and stirred for 15 min. After standing for 30 min, the aqueous layer was removed, and the resulting organic layer was filtered. The filtrate was left standing for 30 min, and the aqueous layer was removed again. The resulting organic solution was solvent-exchanged with isopropanol (5 L / kg, 480 L) and used in the next step without further purification.

[0220] Intermediate 13: 2-[[(1R)-1-(3,6-dimethyl-4-oxo-2-phenyl-chromen-8-yl)ethyl]amino]benzoic acid; (1S)-1-phenylethanamine

[0221]

Chem.

[0222] Compound A: 2-[[(1R)-1-(3,6-dimethyl-4-oxo-2-phenyl-chromen-8-yl)ethyl]amino]benzoic acid

[0223]

Chemical formula

[0224] Route 2: 2-[[(1R)-1-(3,6-dimethyl-4-oxo-2-phenyl-chromen-8-yl)ethyl]amino]benzoic acid; (1S)-1-phenylethylamine (15 g) was mixed with DCM (150 mL). 1 M aqueous HCl (75 mL) was added and the mixture was stirred for 5 min. 1 M aqueous HCl (75 mL) was added and the mixture was stirred again for 5 min. The reaction mixture was concentrated to about 45 mL. DCM (30 mL) and ACN (150 mL) were added and the mixture was concentrated to about 75 mL. ACN (150 mL) was added and the mixture was concentrated to about 120 mL. It was heated briefly at 80 °C. It was cooled to 65 °C and stirred for 2 h. It was cooled to 25 °C over 4 h and stirred for a further 4 h. The product (7.50 g, 65%) was recovered by filtration, washed with 5 mL of ACN, washed with 75 mL of heptane, and dried in a 45 °C oven. MS ES+ m / z 414 [M+H] + .

[0225] Route 3: A flask was charged with 8-[(1S)-1-chloroethyl]-3,6-dimethyl-2-phenyl-chromen-4-one (5.0 g, 16.0 mmol) and IPA (50 mL). The reaction mixture was then treated with anthranilic acid (6.58 g, 48.0 mmol), and triethylamine (6.7 mL, 48.0 mmol) was added dropwise. The reaction mixture was stirred at 60 °C overnight. The reaction mixture was cooled to room temperature, 50 mL of 2-methyltetrahydrofuran was added, and the reaction mixture was concentrated to approximately 50 mL. This solvent exchange was repeated three more times with 25 mL of 2-methyltetrahydrofuran. 25 mL of 2 M HCl aqueous solution was added and stirred. It was transferred to a separatory funnel, and the aqueous layer was removed. The remaining organic layer was washed with 2 M HCl aqueous solution (2 × 25 mL), then diluted with 50 mL of ACN and concentrated three times to approximately 50 mL to leave a thick slurry. The slurry was heated to 80 °C for 1 hour and then at 65 °C for 2 hours. The reaction mixture was cooled to room temperature over about 5 hours and stirred overnight. The product (5.19 g, 79%) was recovered by filtration, washed with 5 mL of ACN, washed with 25 mL of heptane, and dried in an oven at 45 °C.

[0226] Route 4: A solution of 8-[(1S)-1-chloroethyl]-3,6-dimethyl-2-phenyl-chromen-4-one prepared according to an alternative synthesis of Intermediate 12 in isopropanol was charged to a reactor, and anthranilic acid (111.9 kg, 816.3 mol, 2.5 eq) and sodium bicarbonate (41.1 kg, 489.4 mol, 1.5 eq) were added. Isopropanol (2 L / kg, 192 L) was added, and the reaction mixture was stirred at 65 °C for 24 h. Upon completion, the reaction mixture was solvent-exchanged into 2-MeTHF (15 L / kg, 1400 L). At 20 °C, 4 M aqueous HCl solution (5 L / kg, 480 L) was added and stirred for 30 min. After standing for 30 min, the aqueous layer was removed. 2-MeTHF (2 L / kg, 192 L) and 2 M aqueous HCl solution (3 L / kg, 288 L) were added to the organic layer and stirred at 20 °C for 30 min. After standing for 30 min, the aqueous layer was removed. 2-MeTHF (3 L / Kg, 288 L) and water (5 L / kg, 480 L) were added and stirred for 30 min. After standing for 30 min, the aqueous layer was removed. The resulting solution was used in the next step without purification. [Note: Quantities are relative to 8-[(1R)-1-hydroxyethyl]-3,6-dimethyl-2-phenyl-chromen-4-one]

[0227] Alternative Synthesis of Intermediate 13: 2-[[(1R)-1-(3,6-dimethyl-4-oxo-2-phenyl-chromen-8-yl)ethyl]amino]benzoic acid; (1S)-1-phenylethanamine A 2-MeTHF solution of 2-[[(1R)-1-(3,6-dimethyl-4-oxo-2-phenyl-chromen-8-yl)ethyl]amino]benzoic acid was charged into a reactor according to the procedure of Route 4 above and concentrated to 10 L / kg (960 L). The mixture was heated to 50 °C and (S)-methylbenzylamine (6.72 kg, 55.5 mol, 0.17 eq) was added over 1 hour. The resulting slurry was stirred at 50 °C for 45 minutes and additional (S)-methylbenzylamine (303.6 mol, 0.93 eq) was added over 2 hours. The slurry was stirred at 65 °C for 2 hours and then cooled to 20 °C over 12 hours. After stirring at 20 °C for 4 hours, the slurry was filtered and the wet cake was washed with 1:1 2-MeTHF / n-heptane (3 L / kg, 288 L) and n-heptane (3 L / kg, 288 L), and then dried under vacuum at 45 °C for 16 hours to obtain the title compound in 77% yield over 3 steps.

[0228] Example 2 2-[[(1R)-1-(3,6-Dimethyl-4-oxo-2-phenyl-chromen-8-yl)ethyl]amino]benzoic acid Form A (「Compound A Form A」) Compound A Form A was obtained according to the procedure of Example 1.

[0229] The XRPD pattern of Compound A Form A shown in Figure 1 was successfully indexed, confirming that the experimental pattern represented that of a single crystalline phase and that the unit cell volume was consistent with the anhydrous crystalline form.

[0230] XRPD The prepared sample of Form A had diffraction peaks (2-theta values) as described in Table 1 below, and in particular, was characterized by an XRD pattern using CuKα radiation having a peak at 12.1° in combination with one or more of the peaks selected from the group consisting of 14.1°, 16.8°, 18.9° and 20.7°, with an allowable error of 0.2 degrees in diffraction angle.

[0231] Table 1. XRPD Peaks

Table 2

[0232] Thermal analysis Form A was confirmed to be anhydrous based on the lack of mass loss detected upon heating from 25 °C to 200 °C. The sharp endothermic peak at 185 °C was due to the melting of Form A. An exothermic transition was observed at 192 °C, followed by an endothermic event at 211 °C, which corresponded to the crystallization and melting of Form C, respectively.

[0233] Example 3 2-[[(1R)-1-(3,6-Dimethyl-4-oxo-2-phenyl-chromen-8-yl)ethyl]amino]benzoic acid Form B (「Compound A Form B」) A THF solvate was formed by slowly evaporating a saturated THF solution of 2-[[(1R)-1-(3,6-dimethyl-4-oxo-2-phenyl-chromen-8-yl)ethyl]amino]benzoic acid at ambient temperature. The XRPD pattern of the THF solvate filtrate shown in Figure 2 was successfully indexed to confirm that the material was a single crystalline phase named Form B. The unit cell volume was consistent with a mono THF solvate.

[0234] XRPD The prepared sample of Form B had diffraction peaks (2 theta values) as listed in Table 2 below. In particular, it was characterized by an XRD pattern using CuKα radiation having a peak at 9.7° in combination with one or more of the peaks selected from the group consisting of 14.9°, 11.9°, 17.4°, and 7.0°, with an angular tolerance of 0.2 degrees for the diffraction angle.

[0235] Table 2. XRPD Peaks

Table 3

[0236] Thermal analysis When heated at a rate of 10 °C / min, the low temperature endothermic event was due to desolvation, followed by the melting of presumably Compound A Form C at 213 °C. In TGA, the mass loss observed upon heating was consistent with 0.8 molar equivalents of THF.

[0237] Example 4 2-[[(1R)-1-(3,6-Dimethyl-4-oxo-2-phenyl-chromen-8-yl)ethyl]amino]benzoic acid Form C (「Compound A Form C」) Form C was generated by annealing Compound A Form A at 194 °C for 30 minutes. From XRPD indexing (Figure 3), the phase purity of Form C was confirmed, and the unit cell volume was consistent with the anhydrous material.

[0238] XRPD The prepared sample of Form C had diffraction peaks (2-theta values) as described in Table 3 below. In particular, it was characterized by an XRD pattern using CuKα radiation, having a peak at 13.4° in combination with one or more of the peaks selected from the group consisting of 8.5°, 15.7°, 10.6°, and 7.4°, with an angular tolerance of 0.2 degrees for the diffraction angle.

[0239] Table 3. XRPD Peaks

Table 4

[0240] Thermal Analysis The DSC of the isolated Compound A Form C showed an endothermic event at 209 °C due to melting.

[0241] Example 5 2-[[(1R)-1-(3,6-Dimethyl-4-oxo-2-phenyl-chromen-8-yl)ethyl]amino]benzoic acid tromethamine salt Form A (「Compound A tromethamine salt Form A」) The preparation of tromethamine salt form A of compound A was carried out by dissolving 2-[[(1R)-1-(3,6-dimethyl-4-oxo-2-phenyl-chromen-8-yl)ethyl]amino]benzoic acid (2.4 g, 5.80 mmol) and tris(hydroxymethyl)aminomethane (714.5 mg, 5.90 mmol) in 4:1 THF:MeOH (15.9 mL) with stirring at 60 °C and 850 rpm. While stirring, heptane (15.8 mL) was added at 60 °C over 12 hours. The slurry was cooled to 10 °C over 4 hours and then stirred at 10 °C overnight. The solid product was isolated on Whatman paper under vacuum to give the title compound (1.72 g, 88% yield).

[0242] Chiral enhancement The formation and isolation of tromethamine salt form A of compound A provided chiral enhancement of the (R)-enantiomer compared to the free acid (R)-enantiomer content. HPLC of the free acid starting material showed 96.6% (R)-enantiomer and 3.4% (S)-enantiomer. HPLC of the crystalline tromethamine salt showed >99.9% (R)-enantiomer.

[0243] XRPD The prepared sample of the crystalline tromethamine salt has diffraction peaks (2-theta values) as described in Table 4 below, and in particular, is characterized by an XRD pattern (Figure 4) using CuKα radiation, having a peak at 6.4° in combination with one or more of the peaks selected from the group consisting of 8.4°, 10.9°, 16.9° and 22.1°, with an angular tolerance of 0.2 degrees for the diffraction angle.

[0244] Table 4. XRPD peaks

Table 5

[0245] Thermal analysis Compound A tromethamine salt Form A was confirmed to be anhydrous based on a negligible weight loss detected upon heating to 200 °C. DSC analysis showed a small endothermic event at 68 °C (onset) due to the conversion of Form A to Form D. Upon continuous heating, an endothermic event was observed at 203 °C (onset), which coincides with the melting of Form D.

[0246] Single crystal structure analysis An appropriate single crystal was selected and analyzed by single crystal X-ray diffraction. The single crystal structure of Compound A tromethamine salt Form A was determined, and the molecular structure and absolute configuration were confirmed. The structure was determined to be an anhydrous crystal form composed of one Compound A anion and one tromethamine cation in the asymmetric unit. The absolute structure was determined from the crystal structure, and it was found that the molecules are bonded in the R configuration.

[0247] Solid state NMR A prepared sample of Compound A tromethamine salt Form A was characterized by solid state NMR. 13 C Solid state NMR (100.6 MHz) δ 179.0, 158.7, 151.7, 149.7, 136.3, 134.7, 132.9, 129.3, 127.4, 125.2, 121.7, 117.0, 115.5, 115.2, 110.4, 64.1, 63.2, 45.3, 22.6, 20.3, 11.6 ppm.

[0248] Alternative Example 5 2-[[(1R)-1-(3,6-Dimethyl-4-oxo-2-phenyl-chromen-8-yl)ethyl]amino]benzoic acid tromethamine salt Form A (“Compound A tromethamine salt Form A”) The reactor was charged with 2-MeTHF (10 L / kg), 2-[[(1R)-1-(3,6-dimethyl-4-oxo-2-phenyl-chromen-8-yl)ethyl]amino]benzoic acid; (1S)-1-phenylethylamine, and 2N aqueous HCl solution at 25 °C (4 L / kg). The resulting two-phase mixture was stirred for 30 minutes, left standing for 30 minutes, and then the aqueous layer was removed. 2N aqueous HCl solution (4 L / kg) was added to the organic layer, stirred for 30 minutes, and then left standing for 30 minutes. The aqueous layer was removed, and water (4 L / kg) was added to the organic layer. After stirring for 30 minutes, the mixture was allowed to stand for 30 minutes, and the aqueous layer was removed. The resulting organic layer was solvent-exchanged with THF (1.93 L / kg). After adding MeOH (1.93 L / kg), aminotris(hydroxymethyl)methane (0.95 equivalent) was added. The solution was heated to 40 °C, polished filtered (0.22 μm filter), and aged for 30 minutes. n-Heptane (1.29 L / kg) was added, crystallization was seeded (2.5 wt%), and aged for 30 minutes. At 40 °C, n-heptane (3.24 L / kg) was added over 8 hours. A further n-heptane (3.24 L / kg) was added over 4 hours. The resulting slurry was cooled to 10 °C over 5 hours and stirred at 10 °C for 4 hours. The slurry was filtered, and the wet cake was washed with 1:2 (1:1 THF / MeOH):n-heptane (3.8 L / kg), and then with n-heptane (3.8 L / kg). The wet cake was dried under vacuum at 50 °C to obtain the title compound in 87% yield. The solid was pin-milled with a Hosokawa Alpine 160 UPZ pin mill to obtain a material having a particle size of less than 20 micrometers. For the record of the particle size analysis of 4 lots of tromethamine salt form A of 2-[[(1R)-1-(3,6-dimethyl-4-oxo-2-phenyl-chromen-8-yl)ethyl]amino]benzoic acid, see Table 8.

[0249] Table 8. D90 measurement

Table 6

[0250] Example 6 2-[(1R)-1-(3,6-Dimethyl-4-oxo-2-phenyl-chromen-8-yl)ethyl]amino-benzoic acid tromethamine salt form C (“Compound A tromethamine salt form C”) The seed material was prepared by dissolving tris(hydroxymethyl)aminomethane (>1 equivalent) in either water (0.5 - 1.7 mL) and acetone (0.1 - 0.2 mL) or approximately 3:1 water:acetone (0.6 mL), followed by the addition of Compound A form A (30 - 100 mg). If the resulting slurry was difficult to stir, additional acetone was added. The sample was stirred at room temperature for several hours or overnight. The suspension was centrifuged at room temperature for 5 minutes and dried at room temperature overnight to obtain a solid consistent with Compound A tromethamine salt form C as a monohydrate.

[0251] The preparation of Compound A tromethamine salt form C was carried out by combining approximately 3:1 acetone:water (16.5 mL) and Compound A tromethamine salt form A (4.93 g, pulverized). The mixture was stirred at room temperature for 5 minutes (750 rpm). A seed material (monohydrate Compound A tromethamine salt form C, 45 mg) was added to the suspension, and the mixture was stirred at room temperature overnight (750 rpm), and the sample was stirred overnight under the same previous conditions. The suspension was centrifuged at room temperature for 5 minutes to obtain a solid consistent with Compound A tromethamine salt form C as a monohydrate.

[0252] XRPD The prepared sample of Compound A tromethamine salt form C has diffraction peaks (2-theta values) as described in Table 5 below, and in particular, has a peak at 15.9° in combination with one or more of the peaks selected from the group consisting of 10.6°, 17.4°, 13.2°, and 14.5°, and is characterized by an XRD pattern (Figure 5) using CuKα radiation, with an allowable error of 0.2 degrees in diffraction angle.

[0253] Table 5. XRPD Peaks

Table 7

[0254] Thermal analysis In thermal analysis, the desolvation of Form C is indicated by a 3.1% weight loss, which is consistent with the theoretical value of 3.4% for the monohydrate form.

[0255] Example 7 Tromethamine salt of 2-[[(1R)-1-(3,6-dimethyl-4-oxo-2-phenyl-chromen-8-yl)ethyl]amino]benzoic acid, Form D (「Tromethamine salt of Compound A, Form D」) The preparation of the tromethamine salt of Compound A, Form D, was carried out by dispensing the tromethamine salt of Compound A, Form A, into a flat plate XRPD sample holder and heating it to 150 °C. The conversion from Form A to Form D occurred during heating.

[0256] XRPD Form D was only observed in situ (VT-XRPD). Attempts to generate Form D by high-temperature annealing resulted in Form A by XRPD, suggesting that the conversion to Form A occurs within a few minutes at RT.

[0257] The indexing of the XRPD pattern of Form D (Figure 6) collected on VT-XRPD at 150 °C was successful, confirming that the experimental pattern represents that of a single-crystalline phase. The tromethamine salt of Compound A, Form D, has diffraction peaks (2-theta values) as described in Table 6 below, and in particular, is characterized by an XRD pattern using CuKα radiation having a peak at 11.1° in combination with one or more of the peaks selected from the group consisting of 12.6°, 17.1°, 6.3° and 18.9°, with an allowable error of 0.2 degrees in diffraction angle.

[0258] Table 6. XRPD Peaks

Table 8

[0259] Example 8 2-[[(1R)-1-(3,6-Dimethyl-4-oxo-2-phenyl-chromen-8-yl)ethyl]amino]benzoic acid albumin salt Form A (「Compound A albumin salt Form A」) 2-[[(1R)-1-(3,6-Dimethyl-4-oxo-2-phenyl-chromen-8-yl)ethyl]amino]benzoic acid (0.752 g, 1.82 mmol) was suspended in acetone (30 mL) and stirred at 55 °C at 500 rpm. tert-Butylamine (0.220 mL, 2.09 mmol) was added. Acetone (5 mL) was added to dilute the resulting slurry. The slurry was stirred at 55 °C at 450 rpm for 45 minutes, cooled to room temperature, and isolated on Whatman paper under vacuum to obtain the title compound (0.849 g, yield 96%).

[0260] XRPD The prepared sample of the crystalline albumin salt has diffraction peaks (2-theta values) as described in Table 7 below, and in particular, has a peak at 11.1 in combination with one or more of the peaks selected from the group consisting of 10.5, 15.2, 18.0 and 19.3, and is characterized by an XRD pattern (Figure 7) using CuKα radiation, with an allowable error of 0.2 degrees in diffraction angle.

[0261] Table 7. XRPD Peaks

Table 9

[0262] Solid NMR The prepared sample of albumin salt Form A of Compound A was characterized by solid NMR. 13 C Solid state NMR (100.6 MHz) δ 177.4, 174.8, 159.8, 151.7, 149.5, 134.0, 132.6, 130.7, 130.3, 129.0, 128.1, 123.1, 122.4, 119.4, 116.8, 115.9, 112.3, 53.0, 47.5, 27.4, 23.3, 21.3, 11.3 ppm.

[0263] Example 9 PI3K-α kinase (PIK3CA) activity in vitro cell-based assay To measure the inhibitory effect of Compound A on PI3Kα H1047R signaling in cancer cells in vitro, increasing concentrations of the inhibitor were administered to serum-free MDA-MB-453 cells. After 3 hours of treatment, the cells were lysed and phospho-AKT Ser473 was monitored using the SureFire® Ultra® assay (PerkinElmer, catalog number ALSU-PAKT-B50K).

[0264] The MDA-MB-453 (ATCC-HTB-131) cell line was obtained from the American Type Culture Collection (Manassas, VA). The cells were maintained in Dulbecco's Modified Eagle Medium (DMEM, Gibco 11965-092) supplemented with 10% fetal bovine serum, heat-inactivated (FBS HI, Gibco 10082-147), 1× non-essential amino acids (NEAA, Gibco 11140-050), and 1 mM sodium pyruvate (Gibco 11360-070). The cultures were maintained in a humidified incubator at 37 °C under 5% CO2 / 95% air.

[0265] To test the compound in 0% FBS, MDA-MB-453 cells were seeded at 1.5×10 per well into white 384-well plates containing 20 μl of minimum essential medium (MEM) assay medium containing 1× NEAA, 1 mM sodium pyruvate, and 1 μg / mL human insulin (Sigma I9278). 4Seeded at the density of cells. The compounds dissolved in a 10 mM stock solution in DMSO were serially diluted 1:3 in DMSO to create a 10-point dilution series and plated using an acoustic liquid handler system (Echo 550 Series Liquid Handler, Labcyte). Subsequently, a 5× intermediate compound dilution plate in MEM containing 1× NEAA and 1 mM sodium pyruvate (starting compound concentration of 150 μM in 1.5% DMSO) was prepared. 5 μl of the intermediate serial dilution compounds were added to the cell plates to give a final concentration range of 30 mM to 0.0015 mM in 0.3% DMSO. 0.3% DMSO alone was used to establish the maximum (MAX) signal, and GDC-0032 at a final concentration of 1 μM was used as a reference compound for the minimum (MIN) signal. After 3 hours of treatment, the medium was removed and the cells were lysed in 10 μL of 1× SureFire lysis buffer with shaking for 10 minutes at room temperature. The acceptor mix (reaction buffer 1 + reaction buffer 2 + activation buffer + SureFire Ultra acceptor beads) was prepared by diluting the activation buffer 25-fold in the combined reaction buffer 1 and reaction buffer 2. The acceptor beads were diluted 50-fold in the combined reaction buffer. 5 μL of the acceptor mix was added to each well, the plate was sealed, covered with foil, and incubated at room temperature for 1 hour. The donor mix (dilution buffer + SureFire Ultra donor beads) was prepared by diluting the donor beads 50-fold in the dilution buffer. 5 μL of the donor mix was added to each well, the plate was sealed, covered with foil, and incubated in the dark at room temperature for 1 hour. The plate was read on a Biotek Neo2 plate reader device using standard AlphaLisa settings.

[0266] The compounds were tested in duplicate, and a single-dose response curve was created using the average percent inhibition at each compound concentration. The data was processed using the Genedata-Screener tool. Relative IC 50Values were determined by calculating the percent inhibition relative to the "minimum" (see GDC-0032 reference control) and "maximum" (DMSO) controls within the plate using the luminescence units. Data were analyzed using a four-parameter non-linear logistic equation (four-parameter logistic concentration-response curve). Y = Bottom + [(Top - Bottom) / 1 + (X / IC50)Slope] Where Y = % inhibition, X = concentration of inhibitor, Bottom = minimum value of y reached by curve fit, Top = maximum value of y reached by curve fit, and Slope = slope of the curve at IC 50 at the curve. % inhibition = [(Signal at X - Minimum Median) / (Maximum Median - Minimum Median)] × 100 IC 50 : Concentration of a compound that reduces a given response (ligand binding, enzyme response) by 50%. Relative IC 50 : Concentration that gives half of the maximum response of the compound.

[0267] Compound A was determined to have an IC 50 value of 6.83 nanomolar.

Claims

1. The tromethamine salt of 2-[[(1R)-1-(3,6-dimethyl-4-oxo-2-phenyl-chromen-8-yl)ethyl]amino]benzoic acid.

2. The tromethamine salt according to claim 1, characterized by an X-ray powder diffraction pattern using CuKα radiation having at least one peak at diffraction angle 2θ selected from 6.4° ± 0.2°, 8.4° ± 0.2°, 10.9° ± 0.2°, 11.8° ± 0.2°, 13.0° ± 0.2°, 16.5° ± 0.2°, 16.9° ± 0.2°, 22.1° ± 0.2°, 23.0° ± 0.2°, and 24.9° ± 0.2°.

3. The tromethamine salt according to claim 1, characterized by an X-ray powder diffraction pattern using CuKα radiation having a peak at diffraction angle 2θ of 6.4° ± 0.2° together with at least one peak selected from 8.4° ± 0.2°, 10.9° ± 0.2°, 16.9° ± 0.2°, and 22.1° ± 0.2°.

4. The tromethamine salt according to claim 1, characterized by an X-ray powder diffraction pattern using CuKα radiation having a peak at diffraction angle 2θ of 6.4° ± 0.2° together with peaks at 8.4° ± 0.2°, 10.9° ± 0.2°, 16.9° ± 0.2° and 22.1° ± 0.2°.

5. Comprising at least one peak relative to glycine (external standard at 176.5 ppm), selected from 179.0, 158.7, 151.7, 149.7, 136.3, 134.7, 132.9, 129.3, 127.4, 125.2, 121.7, 117.0, 115.5, 115.2, 110.4, 64.1, 63.2, 45.3, 22.6, 20.3, and 11.6 ppm (each ±0.2 ppm). １３ The tromethamine salt according to claim 4, characterized by a solid-state C NMR (100.6 MHz) spectrum.

6. Containing peaks referenced to glycine (external standard at 176.5 ppm) at 179.0, 129.3, 63.2, 20.3, and 11.6 ppm (each ±0.2 ppm) １３ The tromethamine salt according to claim 4, characterized by a solid-state C NMR (100.6 MHz) spectrum

7. The tromethamine salt according to claim 1, characterized by an X-ray powder diffraction pattern using CuKα radiation having at least one peak at diffraction angle 2θ selected from 10.6° ± 0.2°, 13.2° ± 0.2°, 14.5° ± 0.2°, 15.9° ± 0.2°, and 17.4° ± 0.2°.

8. The tromethamine salt according to claim 1, characterized by an X-ray powder diffraction pattern using CuKα radiation having a peak at diffraction angle 2θ of 15.9° ± 0.2° together with at least one peak selected from 10.6° ± 0.2°, 17.4° ± 0.2°, 13.2° ± 0.2°, and 14.5° ± 0.2°.

9. The tromethamine salt according to claim 1, characterized by an X-ray powder diffraction pattern using CuKα radiation having peaks at diffraction angles 2θ of 10.6° ± 0.2°, 13.2° ± 0.2°, 14.5° ± 0.2°, 15.9° ± 0.2° and 17.4° ± 0.2°.

10. The tromethamine salt according to claim 1, characterized by an X-ray powder diffraction pattern using CuKα radiation, having at least one peak at diffraction angle 2θ selected from 6.3° ± 0.2°, 11.1° ± 0.2°, 12.6° ± 0.2°, 17.1° ± 0.2° and 18.9° ± 0.2°.

11. The tromethamine salt according to claim 1, characterized by an X-ray powder diffraction pattern using CuKα radiation, having a peak at diffraction angle 2θ of 11.1° ± 0.2° together with at least one peak selected from 12.6° ± 0.2°, 17.1° ± 0.2°, 6.3° ± 0.2° and 18.9° ± 0.2°.

12. The tromethamine salt according to claim 1, characterized by an X-ray powder diffraction pattern using CuKα radiation, having peaks at diffraction angles 2θ of 6.3° ± 0.2°, 11.1° ± 0.2°, 12.6° ± 0.2°, 17.1° ± 0.2° and 18.9° ± 0.2°.

13. The albumin salt of 2 - [[(1R)-1-(3,6-dimethyl-4-oxo-2-phenyl-chromen-8-yl)ethyl]amino]benzoic acid.

14. The albumin salt according to claim 13, characterized by an X-ray powder diffraction pattern using CuKα radiation, having at least one peak at diffraction angle 2θ selected from 6.5° ± 0.2°, 10.5° ± 0.2°, 11.1° ± 0.2°, 15.2° ± 0.2°, 15.9° ± 0.2°, 17.6° ± 0.2°, 18.0° ± 0.2°, 19.3° ± 0.2°, 21.5° ± 0.2°, 22.2° ± 0.2°, 22.7° ± 0.2° and 26.3° ± 0.2°.

15. The albumin salt according to claim 13, characterized by an X-ray powder diffraction pattern using CuKα radiation, having a peak at diffraction angle 2θ of 11.1° ± 0.2° together with at least one peak selected from 10.5° ± 0.2°, 15.2° ± 0.2°, 18.0° ± 0.2° and 19.3° ± 0.2°.

16. The albumin salt according to claim 13, characterized by an X-ray powder diffraction pattern using CuKα radiation, having a peak at diffraction angle 2θ of 11.1° ± 0.2° together with peaks at 10.5° ± 0.2°, 15.2° ± 0.2°, 18.0° ± 0.2° and 19.3° ± 0.2°.

17. including at least one peak referenced to glycine (external standard at 176.5 ppm) selected from 177.4, 174.8, 159.8, 151.7, 149.5, 134.0, 132.6, 130.7, 130.3, 129.0, 128.1, 123.1, 122.4, 119.4, 116.8, 115.9, 112.3, 53.0, 47.5, 27.4, 23.3, 21.3, and 11.3 ppm (each ±0.2 ppm) １３ The albumin salt according to claim 16, characterized by a solid state C NMR (100.6 MHz) spectrum

18. Containing peaks referenced to glycine (external standard at 176.5 ppm) at 177.4, 132.6, 27.4, 21.3, and 11.3 ppm (±0.2 ppm each). １３ The albumin salt according to claim 16, characterized by a 13C solid-state NMR (100.6 MHz) spectrum.

19. A pharmaceutical composition comprising a tromethamine salt according to any one of claims 1 to 12 or an albumin salt according to any one of claims 13 to 18 and a pharmaceutically acceptable carrier.

20. The pharmaceutical composition according to claim 19, for inhibiting phosphoinositide 3-kinase (PI3K).

21. The pharmaceutical composition according to claim 19, for use in the treatment of a disease associated with mutant phosphoinositide 3-kinase (PI3K).

22. The pharmaceutical composition according to claim 21, wherein the PI3K is PI3Kα.

23. The pharmaceutical composition according to claim 21, wherein the PI3K has an H1047R mutation.

24. The pharmaceutical composition according to claim 21, wherein the disease is cancer.

25. The pharmaceutical composition according to claim 24, wherein the cancer is endometrial cancer, gastric cancer, leukemia, lymphoma, sarcoma, colorectal cancer, lung cancer, ovarian cancer, skin cancer, head and neck cancer, breast cancer, brain cancer, or prostate cancer.

26. The pharmaceutical composition according to claim 24, wherein the cancer is breast cancer.

27. The pharmaceutical composition according to claim 24, wherein the cancer is a hormone receptor positive (HR+), human epidermal growth factor receptor 2 negative (HER2−) advanced or metastatic breast cancer.

28. The pharmaceutical composition according to claim 21, wherein the disease is CLOVES syndrome (Congenital Lipomatous Overgrowth, Vascular Malformations, Epidermal Nevi, Scoliosis / Skeletal and Spinal Syndrome), or PIK3CA-related overgrowth syndrome (PROS).

29. Crystalline 2-[[(1R)-1-(3,6-dimethyl-4-oxo-2-phenyl-chromen-8-yl)ethyl]amino]benzoic acid, characterized by an X-ray powder diffraction pattern using CuKa radiation and having at least one peak at diffraction angles 2 theta selected from 7.0° ± 0.2°, 9.7° ± 0.2°, 11.9° ± 0.2°, 14.9° ± 0.2°, and 17.4° ± 0.2°.

30. Crystalline 2-[[(1R)-1-(3,6-dimethyl-4-oxo-2-phenyl-chromen-8-yl)ethyl]amino]benzoic acid, characterized by an X-ray powder diffraction pattern using CuKa radiation and having at least one peak at diffraction angles 2 theta selected from 7.4° ± 0.2°, 8.5° ± 0.2°, 10.6° ± 0.2°, 13.4° ± 0.2°, and 15.7° ± 0.2.

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